Diagnostic device for detecting the presence of two or more biomarkers

Abstract

A test device, such as a lateral flow device, for detecting the presence of two or more biomarkers selected from the group consisting of fungal, viral and / or bacterial biomarkers, in a liquid sample, wherein the presence of at least one of the biomarkers is detected at two or more different concentrations in the sample. The device is particularly useful in the diagnosis of sepsis and / or the selection of treatment for a subject suspected of having sepsis.

Description

[0001] DIAGNOSTIC DEVICE FOR DETECTING THE PRESENCE OF TWO OR MORE BIOMARKERS

[0002] Field of the Invention

[0003] The present invention relates to diagnostic devices for diagnosis of liquid biological samples (e.g., blood)

[0004] 5 and particularly, although not exclusively, to devices for use in the diagnosis of sepsis.

[0005] Background

[0006] Sepsis is a potentially life-threatening inflammatory condition characterised by a dysregulated immune response to host tissue damage or infection by bacteria, fungi, viruses and, on rare occasions, parasites. Symptoms of sepsis may include a fever, breathing difficulties, increased heart rate and mental confusion. If left untreated, sepsis can progress to septic shock characterised by dangerously low blood pressure and multiple organ dysfunction. It is estimated that currently 20% of all deaths worldwide are sepsis related.

[0007] A number of treatment options are available for the three most common types of pathogen induced5 sepsis; bacterial sepsis may be treated with antibiotics, viral sepsis with antivirals and fungal sepsis with antifungal agents. Epidemiological data varies, but bacterial sepsis may account for as much as 70% of all infective causes of sepsis.

[0008] Fungal infections account for approximately 15% of infections and are associated with some of the highest rates of inappropriate therapy and mortality (Candida species alone accounting for approximately0 5% of all sepsis cases). Viruses account for only a small number of cases, approximately 1% to 2%, but data for viral cases is often not collected. The remaining causes being either mixed co-infections, or of unknown or potentially parasitic origin.

[0009] Hospitalized patients pose a much greater challenge to sepsis prevention because of their concurrent illness and an environment rich in pathogens. Currently, sepsis is initially screened for in healthcare5 settings through periodic clinical observations. In the UK, the National Early Warning Score (NEWS2) assessment system has been implemented by the UK’s national healthcare service which includes analysis of respiratory rate, oxygen saturation, temperature, systolic blood pressure, pulse rate and any cognitive changes (similar screening and assessment protocols exist and are implemented around the world). Should a patient present with abnormal assessment results, sepsis may be suspected and blood samples taken for culture and laboratory evaluation.

[0010] Current blood-based laboratory tests used to aid sepsis diagnosis include blood gas (including glucose and lactate measurements), blood culture, full blood count, C-reactive protein (CRP), urea and electrolytes, creatinine and a clotting screen. Results from blood tests may take a couple of hours and in some cases, a few days from collection, depending on laboratory turnaround times and workflow 5 arrangements. Blood cultures are still seen as the 'gold standard' test for diagnosing sepsis and guiding treatment options, yet approximately 40% of positive blood culture results are false positives due to sample contamination. Furthermore, the results for blood culture tests can take 48 hours for bacterial cultures, with the results from fungal cultures taking as long as 72 hours. It has been suggested that in cases of bacterial sepsis, the chance of survival decreases by approximately 8% for every hour delay before treatment is given, with fungal cases requiring even more urgent treatment.

[0011] Therefore, a first priority in sepsis treatment and improved outcomes is early antimicrobial administration and source control. Current healthcare guidance states that a broad-spectrum antibiotic must be given at the maximum recommended dose without delay (ideally within 1 hour of identifying that the patient meets any high-risk criteria in an acute hospital setting), and not to wait for test results to confirm a bacterial infection. This approach is taken due to the fact that most sepsis cases are caused by a bacterial infection and because the effectiveness of any treatment for sepsis is time critical.

[0012] However, the use of broad-spectrum antibiotics in cases of non-bacterial sepsis can lead to additional complications, such as further opportunistic infections, potential antibiotic resistance and increased morbidity and mortality by delaying more appropriate targeted treatment. Furthermore, the use of unnecessary antibiotics for non-bacterial cases of sepsis is a significant drain on healthcare budgets. The clinical presentation of non-bacterial sepsis is not significantly different from that of bacterial sepsis. For example, fungal sepsis (with rare exceptions) does not present with specific clinical manifestations or laboratory abnormalities, meaning the identification and management of fungal sepsis remains a significant challenge.

[0013] To date, there is no single definitive biomarker for sepsis, that can replace the requirement for additional blood tests. Current point of care diagnostic tests are based on measuring the magnitude of an inflammatory response using non-specific inflammatory biomarkers and as such cannot reliably be used to distinguish between sepsis and other non-infectious critical illnesses, for example; Systemic Inflammatory Response Syndrome (SIRS), which presents with almost identical signs to sepsis. Furthermore, the costs and complexities associated with introducing such point of care test(s) has been a significant barrierto theirwider adoption.

[0014] Therefore, the inventors have identified a need for a reliable, low cost, accessible rapid test device preferably capable of distinguishing between non-infectious critical illnesses and sepsis, which is preferably also capable of broadly identifying the trigger of sepsis as being bacterial, fungal or viral and the severity of such bacterial, fungal or viral infection. This may be used to guide initial treatment and limit the overuse of antibiotics.

[0015] It would be beneficial in the professional setting to have an immediate indication of the presence and severity of non-infectious critical illnesses and / or bacterial, viral and / or fungal triggers of sepsis without the need for a test reader or other electronic type device, which would limit the accessibility of the test in certain clinical settings and add cost and complexity to the test. The present invention has been devised in light of the above considerations. Summary of the Invention

[0016] The invention provides a test device for detecting the presence of biomarkers in a liquid sample, the device comprising: a permeable material defining: a first portion providing a first site for application of the liquid sample and for conjugates movably supported therein wherein each conjugate comprises a binder for a respective biomarker coupled to a respective detector reagent; and, a second portion configured relative to the first portion so as to permit capillary flow communication therebetween, and said second portion providing a second site spaced from the first site for visually determining the presence of the respective detector reagent, and comprising binders immobilized therein each of which binds to a respective said biomarker; wherein the first portion comprises for a first said biomarker, a first said moveably supported conjugate comprising a first binder for a fungal, viral or bacterial biomarker substance coupled to a first respective detector reagent and, for at least one further said biomarker, at least one further said moveably supported conjugate comprising at least one further binder coupled to at least one further detector reagent, wherein the at least one further biomarker comprises a respective biomarker substance selected from a fungal, viral or bacterial biomarker substance, wherein the first biomarker and the at least one further biomarker are different biomarkers, and wherein the second site is configured to visually determine the presence of at least one of the respective detector reagents at two or more different concentrations of at least one of the biomarkers.

[0017] In this way, the test device may provide the combined, simultaneous testing, from the same liquid sample of two or more different biomarkers, for example, both a fungal biomarker and at least one of a viral biomarker and / or a bacterial biomarker, wherein the device can provide a visual indication of two or more concentrations of at least one of the biomarkers present in the sample. In another example, the device may test both a viral biomarker and / or a bacterial biomarker, wherein the device can provide a visual indication of two or more concentrations of at least one of the biomarkers present in the sample.

[0018] The first portion may comprise a first binder for fungal biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site may be configured to visually determine the presence of the fungal biomarker substance at two or more different concentrations of the fungal biomarker substance.

[0019] The first portion may comprise a first binder for a fungal biomarker substance and at least one further binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the bacterial biomarker substance at two or more different concentrations of the bacterial biomarker substance.

[0020] The first portion may comprise a first binder for a fungal biomarker substance and at least one further binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the fungal and bacterial biomarker substance at two or more different concentrations of the fungal and bacterial biomarker substance respectively.

[0021] The first portion may comprise a first binder for viral biomarker substance and at least one further binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the viral biomarker substance at two or more different concentrations of the viral biomarker substance.

[0022] The first portion may comprise a first binder for a viral biomarker substance and at least one further binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the bacterial biomarker substance at two or more different concentrations of the bacterial biomarker substance.

[0023] The first portion may comprise a first binder for a viral biomarker substance and at least one further binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the viral and bacterial biomarker substance at two or more different concentrations of the viral and bacterial biomarker substance respectively.

[0024] The first portion may comprise a first binder for a fungal biomarker substance and at least one further binder for a viral biomarker substance, and the second site may be configured to visually determine the presence of the fungal biomarker substance at two or more different concentrations of the fungal biomarker substance.

[0025] The first portion may comprise a first binder for a fungal biomarker substance and at least one further binder for a viral biomarker substance, and wherein the second site is configured to visually determine the presence of the viral biomarker substance at two or more different concentrations of the viral biomarker substance.

[0026] The first portion may comprise a first binder for a fungal biomarker substance and at least one further binder for a viral biomarker substance, and the second site may be configured to visually determine the presence of the viral biomarker substance and the fungal biomarker substance at two or more different concentrations of the fungal and viral biomarker substance respectively.

[0027] The device may detect the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder binds for a viral biomarker substance and a third binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the fungal biomarker substance at two or more different concentrations of the fungal biomarker substance.

[0028] The device may detect the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the bacterial biomarker substance at two or more different concentrations of the bacterial biomarker substance.

[0029] The device may detect the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the viral biomarker substance at two or more different concentrations of the viral biomarker substance.

[0030] The device may detect the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the fungal and bacterial biomarker substance at two or more different concentrations of the fungal and bacterial biomarker substance respectively.

[0031] The device may detect the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the fungal and viral biomarker substance at two or more different concentrations of the fungal and viral biomarker substance respectively.

[0032] The device may detect the presence of three biomarkers, wherein the first portion comprises a first binder for fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the bacterial and viral biomarker substance at two or more different concentrations of the bacterial and viral biomarker substance respectively.

[0033] The device may detect the presence of three biomarkers, wherein the first portion comprises a binder for a fungal biomarker substance, a second binder binds for a viral biomarker substance and a third binder for a bacterial biomarker substance, and the second site may be configured to visually determine the presence of the fungal, bacterial and viral biomarker substance at two or more different concentrations of the fungal, bacterial and viral biomarker substance respectively.

[0034] The first portion may further comprise a binder for a non-specific inflammatory biomarker substance and the second site may be configured to visually determine the presence of the non-specific inflammatory biomarker at one concentration or at two or more different concentrations of the non-specific inflammatory biomarker substance; optionally wherein the non-specific inflammatory biomarker substance is CRP.

[0035] The fungal biomarker substance may comprise Beta-D-glucan (BDG). The viral biomarker substance may comprise Myxovirus resistance protein A (MxA). The bacterial biomarker substance may comprise Procalcitonin (PCT).

[0036] The second site may comprise two or more test lines comprising binders immobilised therein which bind to at least one respective biomarker, wherein each test line provides a visual determination of the presence of at least one biomarker at two or more different concentrations of said biomarker, optionally, wherein the first test line provides a visual determination of the presence of said at least one biomarker at a first concentration and the second test line provides a visual determination of the presence of said at least one biomarker at a second concentration, optionally, wherein the first test line provides a visual determination of the presence of said at least one biomarker at a low concentration and the second test line provides a visual determination of the presence of said at least one biomarker at a high concentration.

[0037] The first portion may comprise a first binder for a fungal biomarker substance, and the second site may comprise two or more test lines for visually determining the presence of the fungal biomarker in the sample, wherein the fungal biomarker substance may comprise BDG and wherein the first test line may provide a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line may provide a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample.

[0038] The first portion may comprise a first binder for a bacterial biomarker substance, and the second site may comprise two or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance may comprise PCT, and wherein the first test line may provide a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL or below about 0.50 ng / mL in a blood sample and the second test line may provide a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above or about 0.50 ng / mL or above in a blood sample.

[0039] The first portion may comprise a first binder for a bacterial biomarker substance, and the second site may comprise two or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance may comprise PCT, and wherein the first test line may provide a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample and the second test line may provide a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above in a blood sample.

[0040] The first portion may comprise a first binder for a bacterial biomarker substance, and the second site may comprise two or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance may comprise PCT, and wherein the first test line may provide a visual determination of the presence of PCT corresponding to a concentration of below about 0.50 ng / mL in a blood sample and the second test line may provide a visual determination of the presence of PCT corresponding to a concentration of about 0.50 ng / mL or above in a blood sample.

[0041] The first portion may comprise a first binder for a viral biomarker substance, and the second site may comprise two or more test lines for visually determining the presence of the viral biomarker substance in the sample, wherein the viral biomarker substance may comprise MxA and wherein the first test line may provide a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line may provide a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample.

[0042] The first portion may further comprise a binder for a non-specific inflammatory biomarker substance, and the second site may further comprise two or more test lines for visually determining the presence of the non-specific inflammatory biomarker substance in the sample, wherein the non-specific inflammatory biomarker substance may comprise CRP and wherein the first test line may provide a visual determination of the presence of CRP corresponding to a concentration of below about 9.9 mg / dL in a blood sample and the second test line may provide a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

[0043] The second site may comprise three test lines comprising binders immobilised therein which bind to at least one respective biomarker, wherein each test line may provide a visual determination of the presence of said biomarker at three or more different concentrations, wherein the first test line may provide a visual determination of the presence of said at least one biomarker at a first concentration, the second test line may provide a visual determination of the presence of said at least one biomarker at a second concentration, and the third test line may provide a visual determination of the presence of said at least one biomarker at a third concentration, optionally, wherein the first test line provides a visual determination of the presence of said at least one biomarker at a low concentration, the second test line provides a visual determination of the presence of said at least one biomarker at a medium concentration, and the third test line provides a visual determination of the presence of said at least one biomarker at a high concentration.

[0044] The first portion may comprise a first binder for a fungal biomarker substance, and the second site may comprise three or more test lines for visually determining the presence of the fungal biomarker substance in the sample, wherein the fungal biomarker may comprise BDG and wherein the first test line may provide a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample, the second test line may provide a visual determination of BDG corresponding to a concentration of about 81-500 pg / mL in a blood sample and the third test line may provide a visual determination of BDG corresponding to a concentration of above about 500 pg / mL in a blood sample.

[0045] The first portion may comprise a first binder for a viral biomarker substance, and the second site may comprise three or more test lines for visually determining the presence of the viral biomarker substance in the sample, wherein the viral biomarker substance may comprise MxA and wherein the first test line may provide a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample, the second test line may provide a visual determination of the presence of MxA corresponding to a concentration of about 61-100 ng / mL in a blood sample and the third test line may provide a visual determination of the presence of MxA corresponding to a concentration of above about 100 ng / mL in a blood sample. The first portion may comprise a first binder for a bacterial biomarker substance, and the second site may comprise three or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance may comprise PCT, wherein the first test line may provide a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample, the second test line may provide a visual determination of the presence of PCT corresponding to a concentration of about 0.25-0.49 ng / mL in a blood sample and the third test line may provide a visual determination of the presence of PCT corresponding to a concentration of about 0.5 ng / mL or above in a blood sample.

[0046] The first portion may further comprise a binder for a non-specific inflammatory biomarker substance, and the second site may further comprise three or more test lines for visually determining the presence of the non-specific inflammatory biomarker substance in the sample, wherein the non-specific inflammatory biomarker substance may comprise CRP and wherein the first test line may provide a visual determination of the presence of CRP corresponding to a concentration of below about 2.0 mg / dL in a blood sample, the second test line may provide a visual determination of the presence of CRP corresponding to a concentration of about 2.1-9.9 mg / dL in a blood sample and the third test line may provide a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

[0047] The first portion may comprise a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance, a third binder for a bacterial biomarker substance and a fourth binder for a nonspecific inflammatory biomarker substance, and the second site may comprise two or more test lines for visually determining the presence of the fungal, bacterial, viral and non-specific inflammatory biomarker substance in the sample each at two or more different concentrations, wherein the fungal biomarker substance may comprise BDG and wherein the first test line may provide a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line may provide a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, wherein the viral biomarker substance may comprise MxA and wherein the first test line may provide a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line may provide a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample, wherein the bacterial biomarker substance may comprise PCT, and wherein the first test line may provide a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample and the second test line may provide a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above in a blood sample, and wherein the non-specific inflammatory marker may comprise C-reactive protein (CRP), and wherein the first test line may provide a visual determination of the presence of CRP corresponding to a concentration of below about 9.9 mg / dL in a blood sample and the second test line may provide a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample. The first portion may provide comprise a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance, a third binder for a bacterial biomarker substance and a fourth binder for a non-specific inflammatory biomarker substance, and the second site may provide comprise two or more test lines for visually determining the presence of the fungal, bacterial, viral and non-specific inflammatory biomarker substance in the sample each at two or more different concentrations, wherein the fungal biomarker substance may comprise BDG and wherein the first test line may provide a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line may provide a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, wherein the viral biomarker substance may comprise MxA and wherein the first test line may provide a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line may provide a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample, wherein the bacterial biomarker substance may comprise PCT, and wherein the first test line may provide a visual determination of the presence of PCT corresponding to a concentration of below about 0.5 ng / mL in a blood sample and the second test line may provide a visual determination of the presence of PCT corresponding to a concentration of about 0.5 ng / mL or above in a blood sample, and wherein the non-specific inflammatory marker may comprise C-reactive protein (CRP), and wherein the first test line may provide a visual determination of the presence of CRP corresponding to a concentration of below about 9.9 mg / dL in a blood sample and the second test line may provide a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

[0048] The first portion may comprise a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance, a third binder for a bacterial biomarker substance and a fourth binder for a nonspecific inflammatory biomarker substance, and the second site may comprise three or more test lines for visually determining the presence of the fungal, viral, bacterial and non-inflammatory biomarker substances in the sample each at three different concentrations, wherein the fungal biomarker may comprise BDG and wherein the first test line may provide a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample, the second test line may provide a visual determination of BDG corresponding to a concentration of about 81-500 pg / mL in a blood sample and the third test line may provide a visual determination of BDG corresponding to a concentration of above about 500 pg / mL in a blood sample, wherein the viral biomarker substance may comprise MxA and wherein the first test line may provide a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample, the second test line may provide a visual determination of the presence of MxA corresponding to a concentration of about 61-100 ng / mL in a blood sample and the third test line may provide a visual determination of the presence of MxA corresponding to a concentration of above about 100 ng / mL in a blood sample, wherein the bacterial biomarker substance may comprise PCT, wherein the first test line may provide a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample, the second test line may provide a visual determination of the presence of PCT corresponding to a concentration of about 0.25-0.49 ng / mL in a blood sample and the third test line may provide a visual determination of the presence of PCT corresponding to a concentration of about 0.5 ng / mL or above in a blood sample, and wherein the non-specific inflammatory biomarker substance may comprise CRP, wherein the first test line may provide a visual determination of the presence of CRP corresponding to a concentration of below about 2.0 mg / dL in a blood sample, the second test line may provide a visual determination of the presence of CRP corresponding to a concentration of about 2.1-9.9 mg / dL in a blood sample and the third test line may provide a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

[0049] The liquid sample may comprise a blood sample.

[0050] The permeable material may be planar and the second site may be laterally spaced from the first site along the planar permeable material such that the test device provides a lateral flow test device.

[0051] By ‘biomarker’ it may be meant a chemical entity (or substance) that is capable of being measured to detect disease or other biochemical or physiological process and / or responses in a host organism. For example, a biomarker may comprise a naturally occurring molecule, gene, or characteristic by which a particular pathological or physiological process, disease, etc. can be identified. Examples include, but are not limited to: antigens; host derived antibodies; signalling molecules. The term “biomarker”, a portmanteau of “biological marker”, may refer to objective indications of medical state observed from outside the patient which can be measured accurately and reproducibly [see Kyle Strimbu and Jorge A. Tavel, M.D.: Curr Opin HIV AIDS. 2010 November; 5(6): 463-466], The biomarker may be an analyte. The terms analyte and biomarker may be used interchangeably.

[0052] The test device may comprise a further conjugate, and associated immobilized binder, for a non-specific inflammatory biomarker substance. Thus, a yet further biomarker may comprise a non-specific inflammatory biomarker. The non-specific inflammatory biomarker substance may comprise C-Reactive Protein (CRP). Accordingly, the first portion may comprise a conjugate comprising a binder for a nonspecific inflammatory biomarker substance comprising C-Reactive Protein (CRP) coupled to a yet further respective detector reagent, and the second portion may comprise a binder immobilized therein which binds to CRP at a single concentration of CRP. The second portion may comprise a binder immobilized therein which is configured to bind to CRP at two or more different concentrations, e.g. a first and second concentration of CRP, e.g. a low and high concentration of CRP. The second portion may comprise a binder immobilized therein which is configured to bind to CRP at three or more different concentrations, e.g. a first, second and third concentration of CRP, e.g. a low, medium and high concentration of CRP.

[0053] Conjugates for biomarkers, and combinations thereof, may be employed in the test device. The test device may detect two or more biomarkers selected from a bacterial biomarker, a fungal biomarker and a viral biomarker and thus include conjugates for two or more biomarkers selected from a bacterial biomarker, a fungal biomarker and a viral biomarker. For example, the test device may include conjugates for any two or more of: Procalcitonin (PCT), (1-3)-p-d-glucan (BDG), Myxovirus resistance protein A (MxA), Galactomannan (GM). The test device may also include conjugates for C-reactive protein (CRP). The conjugates may include conjugates for PCT and BDG. The conjugates may include conjugates for PCT and MxA. The conjugates may include conjugates for BDG and MxA. The conjugates may include conjugates for PCT, BDG and MxA. The conjugates may include conjugates for GM and PCT. The conjugates may include conjugates for GM and MxA. The conjugates may include conjugates for GM, PCT and MxA. The conjugates may include conjugates for PCT, BDG and CRP. The conjugates may include conjugates for BDG, MxA and CRP. The conjugates may include conjugates for PCT, MxA and CRP. The conjugates may include conjugates for GM, MxA and CRP. The conjugates may include conjugates for GM, PCT and CRP. The conjugates may include conjugates for BDG, PCT, MxA and CRP. The conjugates may include conjugates for GM, PCT, MxA and BDG..

[0054] The test device may detect three biomarkers, wherein the biomarkers are a bacterial biomarker, a fungal biomarker and a viral biomarker and therefore the device may include conjugates for a bacterial biomarker, a fungal biomarker and a viral biomarker. The combination of conjugates for biomarkers included in the test device may comprise conjugates for: BDG and PCT and MxA. The combination of conjugates for biomarkers included in the test device may comprise conjugates for: GM and PCT and MxA. Visual determination of three biomarkers may be provided by use of three immobilized binders respectively for e.g. BDG and PCT and MxA, wherein the test device provides a visual determination of at least one of the biomarkers at two or more concentrations of the respective biomarker. The test device provides a visual determination of at least one of BDG, PCT and MxA at two or more different concentrations of one or more of the biomarkers via two or more test lines for a respective biomarker, wherein the first test line indicates the presence of at least one of BDG, PCT and MxA at a first (e.g. low) concentration and wherein the second test line indicates the presence of the same biomarker at a second (e.g. high) concentration. The device may comprise conjugates for BDG and PCT and MxA, and provide visual determination of BDG and PCT at a single concentration via a single test line each (i.e. one test line of BDG and one test line for PCT) and provide visual determination of MxA at two different concentrations (e.g. low and high concentration) via two test lines for MxA. This configuration provides a test device with at least four test lines. The device may comprise conjugates for BDG and PCT and MxA, and provide visual determination of BDG and PCT at a single concentration via a single test line each (i.e. one test line of BDG and one test line for PCT) and provide visual determination of MxA at three different concentrations (e.g. low, medium and high concentration) via three test lines for MxA. This configuration provides a test device with at least five test lines. The device may comprise conjugates for BDG and PCT and MxA, and provide visual determination of MxA and PCT at a single concentration via a single test line each (i.e. one test line of MxA and one test line for PCT) and provide visual determination of BDG at two different concentrations (e.g. low and high concentration) via two test lines for BDG. This configuration provides a test device with at least four test lines. The device may comprise conjugates for BDG and PCT and MxA, and provide visual determination of MxA and PCT at a single concentration via a single test line each (i.e. one test line of MxA and one test line for PCT) and provide visual determination of BDG at three different concentrations (e.g. low, medium and high concentration) via three test lines for BDG. This configuration provides a test device with at least five test lines. The device may comprise conjugates for BDG and PCT and MxA, and provide visual determination of MxA and BDG at a single concentration via a single test line each (i.e. one test line of MxA and one test line for BDG) and provide visual determination of PCT at two different concentrations (e.g. low and high concentration) via two test lines for PCT. This configuration provides a test device with at least four test lines. The device may comprise conjugates for BDG and PCT and MxA, and provide visual determination of MxA and BDG at a single concentration via a single test line each (i.e. one test line of MxA and one test line for BDG) and provide visual determination of PCT at three different concentrations (e.g. low, medium and high concentration) via three test lines for PCT. This configuration provides a test device with at least five test lines. The test device may further comprise a conjugate for CRP. The test device provides visual determination of CRP at a single concentration (i.e. provide the device with a further test line), two concentrations (e.g. low and high concentrations to provide the device with a further two test lines for CRP) or three concentrations (e.g. low, medium and high concentrations to provide the device with a further three test lines for CRP).

[0055] A combination of conjugates for biomarkers included in the test device may comprise conjugates for: BDG and PCT and MxA and CRP. Visual determination of four biomarkers may be provided by use of four immobilized binders respectively for: BDG and PCT and MxA and CRP, wherein the test device provides a visual determination of at least one of the biomarkers at two or more concentrations of the respective biomarker. The test device may provide visual determination of CRP at a single concentration (i.e. provide the device with a further test line), two concentrations (e.g. low and high concentrations to provide the device with a further two test lines for CRP) or three concentrations (e.g. low, medium and high concentrations to provide the device with a further three test lines for CRP).

[0056] A combination of conjugates for biomarkers included in the test device may comprise conjugates for: GM and PCT and MxA. Visual determination of three biomarkers may be provided by use of three immobilized binders respectively for: GM and PCT and MxA, wherein the test device provides a visual determination of at least one of the biomarkers at two or more concentrations of the respective biomarker. The test device provides a visual determination of at least one of GM, PCT and MxA at two or more different concentrations via two test lines, wherein the first test line indicates the presence of at least one of GM, PCT and MxA at a first (e.g. low) concentration and wherein the second test line indicates the presence of GM, PCT, MxA at a second (e.g. high) concentration. The device may comprise conjugates for GM and PCT and MxA, and provide visual determination of GM and PCT at a single concentration via a single test line each (i.e. one test line of GM and one test line for PCT) and provide visual determination of MxA at two different concentrations (e.g. low and high concentration) via two test lines for MxA. This configuration provides a test device with at least four test lines. The device may comprise conjugates for GM and PCT and MxA, and provide visual determination of GM and PCT at a single concentration via a single test line each (i.e. one test line of GM and one test line for PCT) and provide visual determination of MxA at three different concentrations (e.g. low, medium and high concentration) via three test lines for MxA. This configuration provides a test device with at least five test lines. The device may comprise conjugates for GM and PCT and MxA, and provide visual determination of MxA and PCT at a single concentration via a single test line each (i.e. one test line of MxA and one test line for PCT) and provide visual determination of GM at two different concentrations (e.g. low and high concentration) via two test lines for GM. This configuration provides a test device with at least four test lines. The device may comprise conjugates for GM and PCT and MxA, and provide visual determination of MxA and PCT at a single concentration via a single test line each (i.e. one test line of MxA and one test line for PCT) and provide visual determination of GM at three different concentrations (e.g. low, medium and high concentration) via three test lines for GM. This configuration provides a test device with at least five test lines. The device may comprise conjugates for GM and PCT and MxA, and provide visual determination of MxA and GM at a single concentration via a single test line each (i.e. one test line of MxA and one test line for GM) and provide visual determination of PCT at two different concentrations (e.g. low and high concentration) via two test lines for PCT. This configuration provides a test device with at least four test lines. The device may comprise conjugates for GM and PCT and MxA, and provide visual determination of MxA and GM at a single concentration via a single test line each (i.e. one test line of MxA and one test line for GM) and provide visual determination of PCT at three different concentrations (e.g. low, medium and high concentration) via three test lines for PCT. This configuration provides a test device with at least five test lines. The test device may further comprise a conjugate for CRP. The test device provides visual determination of CRP at a single concentration (i.e. provide the device with a further test line), two concentrations (e.g. low and high concentrations to provide the device with a further two test lines for CRP) or three concentrations (e.g. low, medium and high concentrations to provide the device with a further three test lines for CRP).

[0057] A combination of conjugates for biomarkers included in the test device may comprise conjugates for: GM and PCT and MxA and CRP, wherein the test device provides a visual determination of at least one of the GM, PCT and MxA biomarkers at two or more concentrations of the biomarkers. Visual determination of four biomarkers may be provided by use of four immobilized binders respectively for: GM and PCT and MxA and CRP. The test device may provide visual determination of CRP at a single concentration (i.e. provide the device with a further test line), two concentrations (e.g. low and high concentrations to provide the device with a further two test lines for CRP) or three concentrations (e.g. low, medium and high concentrations to provide the device with a further three test lines for CRP).

[0058] Any of the biomarkers disclosed herein may be measured at a single concentration, two concentrations or more concentrations (e.g. low and high concentration) or three concentrations or more concentrations (low, medium and high concentrations) and can be combined with any of the other biomarkers disclosed herein at a single concentration, two concentrations or more concentrations (e.g. low and high concentration) or three concentrations or more concentrations (low, medium and high concentrations), as outlined in Table 1 .

[0059] The first portion may comprise a porous pad configured for receiving the sample. When the liquid sample is (or comprises) a blood sample, the first portion may remove the red blood cells from the sample (preferably with minimal lysing) so as to enhance the visibility of an accumulation of the detector reagents associated with respective biomarkers.

[0060] For example, the second portion may comprise a porous pad or membrane (e.g., test pad ortest membrane) configured for receiving the sample (or parts of the sample) delivered to it by the first portion. The second porous pad may comprise a membrane of nitrocellulose or similar material. The second porous pad may comprise binders (e.g., immobilised test detection antibodies) for selected biomarkers immobilized at specified regions therein (e.g., to form test “capture lines”) where a user may view the results in the form of a visually detectable presence of the detector reagent (e.g., coloured particles) that have accumulated to form a concentration by being captured there.

[0061] A control site (e.g., a control line) may also be provided at the second portion which comprises a control binder immobilized therein which binds to a control conjugate provided in the first portion, wherein the control conjugate comprises a control detector reagent for visually determining the presence of the control detector reagent at the control site when the control conjugate is bound to the control binder there.

[0062] It is to be understood that the first portion may comprise two separate (but not separated) component parts. The two component parts may divide the first site into two sub-sites that are contiguous with each other or otherwise form a mutual contact supporting a capillary flow of liquid sample between them. A first component part of the first portion (e.g., a first of the two sub-sites) may comprise a porous pad for receiving the liquid sample and for supporting capillary fluid flow of the received sample to a second component part of the first portion (e.g., a second of the two sub-sites) that is in capillary fluid flow communication with the first component part and contains the conjugates movably supported therein. If the liquid sample comprises a blood sample, then the first component part may also serve to remove / filter red blood cells from the sample during capillary flow of the sample thorough it, such that the sample (or at least the remaining parts of it) delivered to the second component part is substantially free of red blood cells. The second component part is typically called a “conjugate pad” in the art, and this may hold the awaiting binders (e.g., binding molecules, such as antibody complexes or the like). Thus, application of the liquid sample may occur at a location upon the porous pad of the first sub-site that is laterally spaced from, and in flow communication with, the location of the conjugates in the porous pad of the second subsite.

[0063] Alternatively, the first portion may comprise one continuous porous pad defining the whole of the first site as opposed to two separate component parts. In other words, one continuous porous pad may provide the site for application of the liquid sample and for conjugates movably supported therein. Of course, application of the liquid sample may occur at a location upon the one continuous porous pad that is laterally spaced from, and in flow communication with, the location of the conjugates.

[0064] Each of the first portion and the second portion may comprise two or more separate sample flow channels collectively defining the first portion and the second portion of the device, wherein: the first portion comprises collectively two (or more) respective channel first portions of the two (or more) sample flow channels (e.g., each formed of respective sample pads and respective conjugate pads etc.) collectively providing the first site (i.e., shared between two (or more) channels) for application of the liquid sample and for a respective conjugate(s) movably supported therein, and the respective conjugate(s) may comprise a binder(s) for a respective biomarker(s) coupled to a respective detector reagent(s) (e.g., coloured particle(s)); and the second portion comprises collectively two (or more) respective channel second portions of the two (or more) channels (e.g., each formed of a respective test zone pad) each being continuous with a respective one of the two (or more) channel first portions and collectively configured relative to a respective one of the two (or more) channel first portions so as to permit capillary flow communication therebetween. This collectively-defined second portion provides the second site in the collective form of two (or more) separate channel second sites (one per channel), each of which is spaced from the collectively-defined first site, for visually determining the presence of a respective detector reagent (e.g., coloured particle) associated with a respective one of the biomarkers. Each one of the two (or more) separate channel second sites comprises a binder(s) immobilized therein which binds to a respective one of the biomarkers, wherein each binder for each biomarker is configured to bind to the biomarker at a single concentration or at two or more concentration (e.g. low / high or low / medium / high) of the respective biomarker. The resulting device structure provides that selected bio marker tests either at a single concentration or at two or more concentration (e.g. low / high or low / medium / high) of the respective biomarker may be separated out into different sample flow channels i.e. certain biomarkers may be tested on either the same sample flow channel as one or more other biomarkers in the device, or separated out to run on their own individual sample flow channel. Separation of certain biomarkers onto individual separate sample flow channels may be desirable in order to increase or decrease the sensitivity of one of the tests, for example, by using different desensitising agents, sensitising agents etc. or to reduce interferences, such as non-specific binding or competition effects.

[0065] The test device may comprise a third portion configured relative to the second portion so as to permit capillary flow communication therebetween for receiving parts of the sample that have passed through the second portion and have not been bound to a binder within the second portion. In this way, the third portion may serve to mop up excess fluid at the end of a test.

[0066] Visually determining the presence of the detector reagent may be done by simple naked eye detection. Visual determination of the detector reagent at the test line confirms the presence of the captured biomarker at the test line corresponding to the selected concentration of that biomarker in a blood sample.

[0067] Detector reagents most preferably comprise particles which, when accumulated in sufficient concentration, visibly display a predefined and recognisable colour. Here the term ‘particles’ may be understood to include a reference to a material in particulate form (e.g., nano-particles of a specified material) as well as a reference to biological particles such as particles (e.g., molecules) of an enzyme substance (e.g., protein molecules). Detector reagents may comprise coloured particles in this sense. Various types of detector reagents can be used for the visualization of a signal. Examples readily available to the person of ordinary skill in the art include (but are not limited to):

[0068] • coloured material in particulate form (e.g., latex beads, colloidal metal particles (e.g., gold), magnetic particles (e.g., permitting additional detection modes using magnetic sensors));

[0069] • particles of a fluorescent material in particulate form (i.e., fluorescing with a pre-defined colour of fluorescent light);

[0070] • enzyme conjugates

[0071] Test-specific binders (binding reagents) may include, without limitation: antibodies, antibody fragments, antigens, enzymes, aptamers or affimers. Any of these may be conjugated to a suitable detection reagent or molecular responder system such as (but not limited to) nanoparticles, nanoshells or enzyme linked systems for the selective binding and detection of each individual biomarker. Examples of suitable binders (e.g. antibodies, etc.) that may be used as detector and / or responder antibodies are (but not limited to) the following:

[0072] CRP Antibodies: Available from BBI Solutions; CRP Recombinant Fab Monoclonal Antibody BR228-D4A3. Available from Creative Biolabs; Anti-CRP Recombinant Antibody (clone E8-G1) (CAT#: VS3-WK63).

[0073] PCT Antibodies: Available from BBI Solutions; Anti- Procalcitonin BM448-V4A1 , BM448-K8C7, BM448-H9F1.

[0074] BDG Antibodies: Available from Creative Biolabs; Recombinant Anti-1 ,3-Beta-glucan Antibody (CAT#: MOB-0228MC). Available from ThermoFisher Scientific; Fungal beta glucan Monoclonal Antibody (8201), Invitrogen™.

[0075] MxA Antibodies: Available from NSJ Bioreagents; Myxovirus resistance protein 1 Antibody / MX1 / MxA (RQ7192). Available from Sigma Aldrich; Anti-MxA, clone M143 (CL143).

[0076] GM Antibodies: Available from ThermoFisher Scientific; Aspergillus Monoclonal Antibody (5145). Available from Creative Biolabs; Mouse Anti-Galactomannan-2 Monoclonal Antibody (CGYJ217).

[0077] The binder may bind to a specific biomarker either directly i.e. through direct interaction with the surface of the target biomarker, or alternatively, the binder may bind to the target biomarker indirectly i.e. without direct contact with the surface of the target biomarker, for example; via another substance, reagent, binder, or molecular system which is interacting with the target biomarker.

[0078] Preferably, the detector reagent, e.g., when a material in a particulate form, comprises a population of particles that is monodisperse (e.g., characterized by particles of uniform size in a dispersed phase). For example, particles of the detector reagent comprise a substantially spherical shape. In use, when a test is run using the test device, the particles are required to move through a torturous pore structure of the first and second portions of the test device. Particles (e.g., material in particulate form) may comprise an average diameter between 5nm-150nm, even more preferably still, a diameter between 20nm-80nm.

[0079] The first biomarker may comprise a fungal biomarker substance, a viral biomarker substance or a bacterial biomarker substance. If the first biomarker comprises a fungal biomarker substance, then the at least one further biomarker may comprise only a viral biomarker substance, or only a bacterial biomarker substance (i.e. the first biomarker and the second biomarker are different). The at least one further biomarker may comprise both (i.e., two separate biomarkers present simultaneously) a viral biomarker substance, and a bacterial biomarker substance. If the first biomarker comprises a viral biomarker substance, then the at least one further biomarker may comprise only a bacterial biomarker substance, or only a fungal biomarker substance. The at least one further biomarker may comprise both a bacterial biomarker substance and a fungal biomarker substance. If the first biomarker comprises a bacterial biomarker substance, then the at least one further biomarker may comprise only a viral biomarker substance, or only a fungal biomarker substance. The at least one further biomarker may comprise both a viral biomarker substance and a fungal biomarker substance. At least one of the biomarkers is tested at two or more different concentrations (e.g. a high and low concentration, or a high, medium and low concentration).

[0080] Accordingly, the test device may be used to detect a fungal biomarker and one more biomarker which is a viral biomarker, wherein the fungal biomarker is tested at two or more different concentrations e.g. employing three test lines in total, one test line for the viral biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the fungal biomarker and one test line for a second concentration (e.g. high concentration of the fungal biomarker). The test device may be used to detect a fungal biomarker and one more biomarker which could just be a viral biomarker, wherein the viral biomarker is tested at two or more different concentrations e.g. employing three test lines in total, one test line for the fungal biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the viral biomarker and one test line for a second concentration (e.g. high concentration) of the viral biomarker. The test device may be used to detect a fungal biomarker and one more biomarker which is a viral biomarker, wherein the viral and fungal biomarkers are both tested at two or more different concentrations e.g. employing four test lines in total, one test line for a first concentration (e.g. a low concentration) of the viral biomarker, one test line for a second concentration (e.g. a high concentration) of the viral biomarker, one test line for a first concentration (e.g. a low concentration) of the fungal biomarker and one test line for a second concentration (e g. high concentration of the fungal biomarker).

[0081] The test device may be used to detect a fungal biomarker and one more biomarker which is a bacterial biomarker, wherein the fungal biomarker is tested at two or more different concentrations e.g. employing three test lines in total, one test line for the bacterial biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the fungal biomarker and one test line for a second concentration (e.g. high concentration) of the fungal biomarker. The test device may be used to detect a fungal biomarker and one more biomarker which could just be a bacterial biomarker, wherein the bacterial biomarker is tested at two or more different concentrations e.g. employing three test lines in total, one test line for the fungal biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the bacterial biomarker and one test line for a second concentration (e.g. high concentration) of the bacterial biomarker. The test device may be used to detect a fungal biomarker and one more biomarker which is a bacterial biomarker, wherein the bacterial and fungal biomarkers are both tested at two or more different concentrations e.g. employing four test lines in total, one test line for a first concentration (e.g. a low concentration) of the bacterial biomarker, one test line for a second concentration (e.g. a high concentration) of the bacterial biomarker, one test line for a first concentration (e.g. a low concentration) of the fungal biomarker and one test line for a second concentration (e.g. high concentration of the fungal biomarker).

[0082] The test device may be used to detect a viral biomarker and one more biomarker which is a bacterial biomarker, wherein the viral biomarker is tested at two or more different concentrations e.g. employing three test lines in total, one test line for the bacterial biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the viral biomarker and one test line for a second concentration (e.g. high concentration) of the viral biomarker. The test device may be used to detect a viral biomarker and one more biomarker which could just be a bacterial biomarker, wherein the bacterial biomarker is tested at two or more different concentrations e.g. employing three test lines in total, one test line for the viral biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the bacterial biomarker and one test line for a second concentration (e.g. high concentration) of the bacterial biomarker. The test device may be used to detect a viral biomarker and one more biomarker which is a bacterial biomarker, wherein the viral and bacterial biomarkers are both tested at two or more different concentrations e.g. employing four test lines in total, one test line for a first concentration (e.g. a low concentration) of the viral biomarker, one test line for a second concentration (e.g. a high concentration) of the viral biomarker, one test line for a first concentration (e.g. a low concentration) of the bacterial biomarker and one test line for a second concentration (e.g. high concentration of the bacterial biomarker).

[0083] The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the fungal biomarker is tested at two or more different concentrations e.g. employing four test lines in total, one test line for the bacterial biomarker at a single concentration, one test line for the viral biomarker at a single concentration, one test line for the fungal biomarker at a first concentration (e.g. a low concentration) of the fungal biomarker and one test line for a second concentration (e.g. high concentration) of the fungal biomarker. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the bacterial biomarker is tested at two or more different concentrations e.g. employing four test lines in total, one test line for the fungal biomarker at a single concentration, one test line for the viral biomarker at a single concentration, one test line for the bacterial biomarker at a first concentration (e.g. a low concentration) of the bacterial biomarker and one test line for a second concentration (e.g. high concentration) of the bacterial biomarker. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the viral biomarker is tested at two or more different concentrations e.g. employing four test lines in total, one test line for the fungal biomarker at a single concentration, one test line for the bacterial biomarker at a single concentration, one test line for the viral biomarker at a first concentration (e.g. a low concentration) of the viral biomarker and one test line for a second concentration (e.g. high concentration) of the viral biomarker. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the fungal and viral biomarkers are tested at two or more different concentrations e.g. employing five test lines in total, one test line for the fungal biomarker at a first concentration (e.g. low concentration) of the fungal biomarker, one test line for the fungal biomarker at a second concentration (e.g. high concentration) of the fungal biomarker, one test line for the viral biomarker at a first concentration (e.g. a low concentration) of the viral biomarker and one test line for a second concentration (e.g. high concentration) of the viral biomarker, and one test line for the bacterial biomarker at a single concentration. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the fungal and bacterial biomarkers are tested at two or more different concentrations e.g. employing five test lines in total, one test line for the fungal biomarker at a first concentration (e.g. low concentration) of the fungal biomarker, one test line for the fungal biomarker at a second concentration (e.g. high concentration) of the fungal biomarker, one test line for the bacterial biomarker at a first concentration (e.g. a low concentration) of the bacterial biomarker and one test line for a second concentration (e.g. high concentration) of the bacterial biomarker, and one test line forthe viral biomarker at a single concentration. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the viral and bacterial biomarkers are tested at two or more different concentrations e.g. employing five test lines in total, one test line for the viral biomarker at a first concentration (e.g. low concentration) of the viral biomarker, one test line forthe viral biomarker at a second concentration (e.g. high concentration) of the viral biomarker, one test line for the bacterial biomarker at a first concentration (e.g. a low concentration) of the bacterial biomarker and one test line for a second concentration (e.g. high concentration) of the bacterial biomarker, and one test line for the fungal biomarker at a single concentration. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the viral, fungal and bacterial biomarkers are tested at two or more different concentrations e.g. employing six test lines in total, one test line for the viral biomarker at a first concentration (e.g. low concentration) of the viral biomarker, one test line for the viral biomarker at a second concentration (e.g. high concentration) of the viral biomarker, one test line for the bacterial biomarker at a first concentration (e.g. a low concentration) of the bacterial biomarker, one test line for a second concentration (e.g. high concentration) of the bacterial biomarker, and one test line for the fungal biomarker at a first concentration (e.g. a low concentration) of the fungal biomarker, one test line for a second concentration (e.g. high concentration) of the fungal biomarker.

[0084] The test device may be used to detect a fungal biomarker and one more biomarker which is a viral biomarker, wherein the fungal biomarker is tested at three or more different concentrations e.g. employing four test lines in total, one test line for the viral biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the fungal biomarker, one test line for a second concentration (e.g. medium concentration) of the fungal biomarker and one test line for a third concentration (e.g. high concentration) of the fungal biomarker. The test device may be used to detect a fungal biomarker and one more biomarker which could just be a viral biomarker, wherein the viral biomarker is tested at three or more different concentrations e.g. employing three test lines in total, one test line for the fungal biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the viral biomarker, one test line for a second concentration (e.g. medium concentration) of the viral biomarker and one line for a third concentration (e.g. high concentration) of the viral biomarker. The test device may be used to detect a fungal biomarker and one more biomarker which could be a viral biomarker, wherein the viral and fungal biomarker are both tested at three or more different concentrations each e.g. employing six test lines in total, one test line for a first concentration (e.g. a low concentration) of the fungal biomarker, one test line for a second concentration (e.g. medium concentration) of the fungal biomarker and one line for a third concentration (e.g. high concentration) of the fungal biomarker, one test line for a first concentration (e.g. a low concentration) of the viral biomarker, one test line for a second concentration (e.g. medium concentration) of the viral biomarker and one line for a third concentration (e.g. high concentration) of the viral biomarker. The test device may be used to detect a fungal biomarker and one more biomarker which is a bacterial biomarker, wherein the fungal biomarker is tested at three or more different concentrations e.g. employing four test lines in total, one test line for the bacterial biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the fungal biomarker, one test line for a second concentration (e.g. medium concentration of the fungal biomarker and one test line for a third concentration (e.g. high concentration) of the fungal biomarker). The test device may be used to detect a fungal biomarker and one more biomarker which could just be a bacterial biomarker, wherein the bacterial biomarker is tested at three or more different concentrations (e.g. employing four test lines in total, one test line for the fungal biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the bacterial biomarker, one test line for a second concentration (e.g. medium concentration) of the bacterial biomarker and one test line for a third concentration (e.g. high concentration) of the bacterial biomarker. The test device may be used to detect a fungal biomarker and one more biomarker which could be a bacterial biomarker, wherein the bacterial and fungal biomarker are both tested at three or more different concentrations each e.g. employing six test lines in total, one test line for a first concentration (e.g. a low concentration) of the fungal biomarker, one test line for a second concentration (e.g. medium concentration) of the fungal biomarker and one line for a third concentration (e.g. high concentration) of the fungal biomarker, one test line for a first concentration (e.g. a low concentration) of the bacterial biomarker, one test line for a second concentration (e.g. medium concentration) of the bacterial biomarker and one line for a third concentration (e.g. high concentration) of the bacterial biomarker. The test device may be used to detect a viral biomarker and one more biomarker which is a bacterial biomarker, wherein the viral biomarker is tested at three or more different concentrations e.g. employing four test lines in total, one test line for the bacterial biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the viral biomarker, one test line for a second concentration (e.g. medium concentration) of the viral biomarker and one test line for a third concentration (e.g. high concentration) of the viral biomarker. The test device may be used to detect a viral biomarker and one more biomarker which could just be a bacterial biomarker, wherein the bacterial biomarker is tested at three or more different concentrations e.g. employing four test lines in total, one test line for the viral biomarker at a single concentration, one test line for a first concentration (e.g. a low concentration) of the bacterial biomarker, one test line for a second concentration (e.g. medium concentration) of the bacterial biomarker and a third test line for a third concentration (e.g. high concentration) of the bacterial biomarker. The test device may be used to detect a viral biomarker and one more biomarker which could be a bacterial biomarker, wherein the bacterial and viral biomarker are both tested at three or more different concentrations each e.g. employing six test lines in total, one test line for a first concentration (e.g. a low concentration) of the viral biomarker, one test line for a second concentration (e.g. medium concentration) of the viral biomarker and one line for a third concentration (e.g. high concentration) of the viral biomarker, one test line for a first concentration (e.g. a low concentration) of the bacterial biomarker, one test line for a second concentration (e.g. medium concentration) of the bacterial biomarker and one line for a third concentration (e.g. high concentration) of the bacterial biomarker. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the fungal biomarker is tested at three or more different concentrations e.g. employing five test lines in total, one test line for the bacterial biomarker at a single concentration, one test line for the viral biomarker at a single concentration, one test line for the fungal biomarker at a first concentration (e.g. a low concentration) of the fungal biomarker, one test line for a second concentration (e.g. medium concentration) of the fungal biomarker and one test line for a third concentration (e.g. high concentration) of the fungal biomarker. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the bacterial biomarker is tested at three or more different concentrations e.g. employing five test lines in total, one test line for the fungal biomarker at a single concentration, one test line for the viral biomarker at a single concentration, one test line for the bacterial biomarker at a first concentration (e.g. a low concentration) of the bacterial biomarker, one test line for a second concentration (e.g. medium concentration) of the bacterial biomarker and one test line for a third concentration (e.g. high concentration) of the bacterial biomarker. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the viral biomarker is tested at three or more different concentrations e.g. employing five test lines in total, one test line for the fungal biomarker at a single concentration, one test line forthe bacterial biomarker at a single concentration, one test line for the viral biomarker at a first concentration (e.g. a low concentration) of the viral biomarker, one test line for a second concentration (e.g. medium concentration) and one test line for a third concentration (e.g. high concentration) of the viral biomarker. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the fungal and bacterial biomarkers are tested at three or more different concentrations, e.g. employing seven test lines in total, one test line for the viral biomarker at a single concentration, one test line for the bacterial biomarker at a first concentration (e.g. a low concentration) of the bacterial biomarker, one test line for the bacterial biomarker at a second concentration (e g a medium concentration), one test line for the bacterial biomarker at a third concentration (e.g. a high concentration), one test line forthe fungal biomarker at a first concentration (e.g. a low concentration) of the fungal biomarker, one test line for a second concentration (e.g. medium concentration) of the fungal biomarker and one test line for a third concentration (e.g. high concentration) of the fungal biomarker. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the fungal and viral biomarkers are tested at three or more different concentrations, e.g. employing seven test lines in total, one test line for the bacterial biomarker at a single concentration, one test line for the viral biomarker at a first concentration (e.g. a low concentration) of the viral biomarker, one test line forthe viral biomarker at a second concentration (e.g a medium concentration), one test line for the viral biomarker at a third concentration (e.g. a high concentration), one test line forthe fungal biomarker at a first concentration (e.g. a low concentration) of the fungal biomarker, one test line for a second concentration (e.g. medium concentration) of the fungal biomarker and one test line for a third concentration (e.g. high concentration) of the fungal biomarker. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the bacterial and viral biomarkers are tested at three or more different concentrations, e.g. employing seven test lines in total, one test line for the fungal biomarker at a single concentration, one test line for the viral biomarker at a first concentration (e.g. a low concentration) of the viral biomarker, one test line for the viral biomarker at a second concentration (e.g a medium concentration), one test line for the viral biomarker at a third concentration (e.g. a high concentration), one test line forthe bacterial biomarker at a first concentration (e.g. a low concentration) of the bacterial biomarker, one test line for a second concentration (e.g. medium concentration) of the bacterial biomarker and one test line for a third concentration (e.g. high concentration) of the bacterial biomarker. The test device may be used to detect a fungal biomarker and two more biomarkers which could include a viral biomarker and a bacterial biomarker, wherein the bacterial, fungal and viral biomarkers are tested at three or more different concentrations each, e.g. employing nine test lines in total, one test line for the fungal biomarker at a first concentration (e.g. a low concentration) of the fungal biomarker, one test line for the fungal biomarker at a second concentration (e.g a medium concentration), one test line for the fungal biomarker at a third concentration (e.g. a high concentration), one test line for the viral biomarker at a first concentration (e.g. a low concentration) of the viral biomarker, one test line for the viral biomarker at a second concentration (e.g a medium concentration), one test line for the viral biomarker at a third concentration (e.g. a high concentration), one test line for the bacterial biomarker at a first concentration (e.g. a low concentration) of the bacterial biomarker, one test line for a second concentration (e.g. medium concentration) of the bacterial biomarker and one test line for a third concentration (e.g. high concentration) of the bacterial biomarker.

[0085] The test device may comprise two conjugates in total, e.g. the first being fungal and the second being either bacterial or viral, or the first being bacterial and the second being viral, or may comprise three conjugates in total, e.g. the first being fungal, the second being bacterial and the third being viral. Indeed, the test device may comprise four conjugates in total, the first being fungal, the second being bacterial, the third being viral and the fourth being a conjugate for a non-specific inflammatory biomarker (e.g. CRP, as discussed below). At least one of the fungal, viral or bacterial biomarkers are detected at two or more different concentrations, e.g. via two or more test lines.

[0086] Most preferably, the colour of the detector reagents (e.g., coloured particles) is visually distinct from the colour of the second portion such that a concentration of immobilized detector reagents (e.g., coloured particles) at the second site is visually identifiable / determinable by the user as a concentration of a colour, in use, distinct from the colour of the parts of the second portion that surround (or are immediately adjacent to) the second site. The second site may be a linear section of the second portion, such that an accumulation of detector reagents (e.g., coloured particles) forms a visible coloured line upon the second portion. For example, the second site may be a test line defined by the linear section of the test pad or test membrane defined by the second portion. Preferably, test lines for respective analytes are separated from each other laterally along the second portion. The colour of any one of the detector reagents (e.g., coloured particles) may be the same as, or different from, the colour of any one of (or each of) the other detector reagents. The test lines for each biomarker (or set of test lines at two or more concentrations) are specific to an individual biomarker by the selection of a specific immobilized binder, i.e. test lines or sets of test lines are not shared between different biomarkers.

[0087] The test device detects at least two biomarkers selected from: a fungal biomarker, a viral biomarker, a bacterial biomarker and any combination thereof. The test device detects at least one biomarker at two or more concentrations (e.g. a low and high concentration, or a low, medium and high concentration). The test device may also detect a non-specific inflammatory biomarker at a single concentration or at two or more concentrations (e.g. a low and high concentration, or a low, medium and high concentration) in combination with any of the other biomarkers disclosed herein. The test device disclosed herein can therefore provide an instant indication of the presence and the severity of a particular infection type (bacterial, fungal and / or viral), and the presence and severity of any general non-specific inflammation, through interpretation of the various test line results for each biomarker allowing clinicians to rapidly select the most appropriate form of treatment for their patient. The test device may be employed at the point of need in any healthcare setting for patients who arrive presenting sepsis symptoms, allowing for more rapid admission and targeted treatment. The invention, in preferred aspects, may be implemented as a lateral flow device (LFD), also known as a lateral flow test (LFT), lateral flow immunoassay (LFIA) or lateral flow assay (LFA). This may provide a simple, low cost, rapid diagnostic solution for the initial diagnosis of a range of diseases.

[0088] Such a test need not (but could) replace the use of lab-based tests and cultures, rather, such a test could be used to establish a more rapid confirmation of the correct treatment course forthe patient at a significantly lower cost than current test methods. This, may help in significantly increase survival rates, reduce hospital stay times, reduce the overall cost of treatment and enhance antibiotic stewardship measures.

[0089] A test device which can indicate whether a particular biomarker is present in the sample at a particular concentration level (e.g. to indicate a high or low concentration, or a high, low or medium concentration of a particular biomarker within a sample) is advantageous in a clinical setting, particularly when used on a sample e.g. obtained from a patient who is suspected of having sepsis. It allows clinicians to rapidly confirm the presence and also the severity of the fungal and / or bacterial and / or viral infection (e.g. the fungal and / or bacterial and / or viral infection causing sepsis). If a non-specific inflammatory biomarker is also detected in the device either at a single concentration or at two or more concentrations, then the device can also detect the presence and severity of inflammation in the sample. This aids the level and urgency of a clinical response and allows the clinicians to rapidly determine the most appropriate treatment option. The test device can also provide valuable prognostic information. The test device can also be used to monitor the effectiveness of any treatment and aid in the safe termination of said treatment.

[0090] Examples of nonspecific inflammatory biomarkers showing widely accepted correlation with sepsis cases include IL-6, IL-10 and CRP. Preferably, the non-specific inflammatory biomarker is CRP.

[0091] Procalcitonin (PCT) is capable of distinguishing bacterial sepsis from other non-infectious critical illness and to guide the use of antibiotic therapy. In healthy individuals, procalcitonin levels are nearly undetectable, rising significantly in a response to a pro-inflammatory stimulus, especially of bacterial origin. Furthermore, it does not rise significantly with viral or non-infectious inflammation.

[0092] Likewise, (1-3)-p-d-glucan (BDG) may be used to detect the presence of fungal infections. BDG is an attractive biomarker as it originates from the cell walls of a broad range of fungal agents, including the commonly encountered agents Candida spp., Aspergillus spp., and Pneumocystis jirovecii.

[0093] Current turnaround times for PCT blood tests in hospital laboratories is approximately 3 to 5 hours, which is a long time to wait in the context of sepsis diagnosis, and the turnaround time for BDG blood tests in some hospital laboratories is approximately 48 hours which is a dangerously long time in the context of sepsis diagnosis. In view of this, the invention may provide an improvement in how data obtainable from existing accepted biomarkers is made available quickly and easily at the actual point of need (e.g., at the bedside) to promptly guide earlier and more effective treatment. The liquid sample may be (or comprise) a blood sample. For example, the sample may be a ‘raw’ and undiluted blood sample, or may be a liquid sample comprising blood and further comprising a diluent such as a test diluent, of pre-treatment. For example, a liquid sample may comprise a diluent comprising a test enhancing reagent(s) such as specific lysing agent(s) (e.g., this may be a preferred option when using for MxA, as discussed herein) or may comprise agglutinating agents, lysing agents which may be added to dilute a ‘raw’ blood sample prior to the sample being administered to the test device or while in the test device. For example, a user of the device one may wish to agglutinate the red blood cells in a diluent first, and / or lyse white blood cells to release any additional intracellular MxA in a diluent pot, and to use the resulting diluted blood sample ad the liquid sample to apply to the test device.

[0094] A concentration of an immobilized binder, for a given biomarker, within the second portion may be provided such that an accumulation of a detector reagent there is (e.g., becomes visible) visually determinable as bound to the immobilized binder (e.g., to form a visible test line result), or such that an accumulation of a detector reagent saturates, when a concentration of the given biomarker within the liquid sample has a single pre-set threshold (e.g. concentration) value or two or more pre-set threshold (e.g. high / low or high / medium / low concentration) value. Preferably, the pre-set threshold values in question is set relative to an equivalent concentration of the biomarker in a corresponding sample of ‘raw’ undiluted blood. This allow users of the test device to employ test diluents, pre-treatments etc, without affecting the clinical concentrations they wish to use for diagnosis i.e., the test line concentration points are preferably set relative to the undiluted clinical threshold value for each biomarker in question. In other words, the pre-set threshold values may be considered to be a minimum sensitivity value for biomarker detection, in terms of concentration within the liquid sample, which is applicable to ‘raw’, undiluted blood samples having by definition a higher biomarker concentration than they would otherwise have if diluted by a diluent etc.

[0095] For example, if the liquid sample is intended to be a diluted blood sample comprising a diluent added to a ’raw’ blood sample that dilutes the ‘raw’ blood sample by a pre-set dilution degree / amount, then the concentration of an immobilized binder, for a given biomarker, within the second portion may be provided such that an accumulation of a detector reagent there is (e.g., becomes visible) visually determinable as bound to the immobilized binder (e.g., to form a visible test line result), or such that an accumulation of a detector reagent saturates, when a concentration of the given biomarker within the diluted blood sample has a pre-set threshold value that corresponds directly to (i.e., is equivalent to, or is an objective proxy for) a pre-set concentration of the given biomarker within the ‘raw’, undiluted blood sample in accordance with the pre-set dilution degree / amount. In those circumstances, when the device is configured to be used on diluted blood samples the configuration of an immobilized binder (e.g., concentration of the binder, or sensitivity of the binder) for a given biomarker within the second portion (or the visibility of the detector reagent) may be adjusted (e.g., be higher) relative to what it would be the case when the test device is configured to be used on ‘raw’, undiluted samples, but nevertheless both configurations (‘raw’ use and ‘diluted’ use) are usable to detect the same pre-set concentration of biomarker within a liquid sample comprising blood that they are configured to diagnose. Throughout this application, the term “about” is used to indicate that a given value may include a value more or less than 10% of the recited value. Alternatively, the given value does not include a value more and less than 10% of the recited value.

[0096] In the test device, according to some embodiments of the invention, the fungal biomarker substance may comprise Beta-D-glucan (BDG). BDG may be detected at a single concentration, or at two or more concentrations (e.g. a low and high concentration, or a low, medium and high concentration).

[0097] When the device is configured to detect BDG at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) is visually determinable as bound to the immobilized binder via the fungal biomarker comprising BDG, when a concentration of the fungal biomarker within the liquid sample corresponds to at least about 60pg / mL in an (e.g., undiluted) blood sample. The inventors have found that this concentration of BDG biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to start to be reliably visually determined. The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result.

[0098] When the device is configured to detect BDG at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that n accumulation of the first detector reagent (e.g., coloured particles) bound to the immobilized binder via the fungal biomarker comprising BDG saturates when a concentration of the fungal biomarker within the liquid sample corresponds to at least about 180pg / mL in an (e.g., undiluted) blood sample. The inventors have found that this concentration of BDG biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to create a maximally visually determinable accumulation of detector reagent (e.g., coloured particles). The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result.

[0099] When the device is configured so that BDG is detected at two or more concentrations, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the fungal biomarker comprising BDG may saturate at a first test line when a concentration of the fungal biomarker within the liquid sample corresponds to below about 80 pg / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the fungal biomarker within the liquid sample corresponds to above about 80 pg / mL (e.g. about 81 pg / mL or above) in an (e.g., undiluted) blood sample. The accumulation of the detector reagent (e.g. coloured particles) bound to the immobilized binder via the fungal biomarker comprising BDG may saturate at a first test line when a concentration of the fungal biomarker within the liquid sample corresponds to between about 60- 80 pg / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the fungal biomarker within the liquid sample corresponds to about 80 pg / mL (e.g. about 81 pg / mL or above) in an (e.g., undiluted) blood sample. When the device is configured so that BDG is detected at three or more concentrations, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the fungal biomarker comprising BDG may saturate at a first test line when a concentration of the fungal biomarker within the liquid sample corresponds to below about 80 pg / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the fungal biomarker within the liquid sample corresponds to between about 81-500 pg / mL in an (e.g., undiluted) blood sample and may saturate a third test line when a concentration of the fungal biomarker within the liquid sample corresponds to above about 500 pg / mL (i.e. about 501 pg / mL or above) in an (e.g., undiluted) blood sample. The accumulation of the detector reagent (e.g. coloured particles) bound to the immobilized binder via the fungal biomarker comprising BDG may saturate at a first test line when a concentration of the fungal biomarker within the liquid sample corresponds to between about 60-80 pg / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the fungal biomarker within the liquid sample corresponds to about 81-500 pg / mL in an (e.g., undiluted) blood sample and may saturate at a third test line when a concentration of the fungal biomarker within the liquid sample corresponds to above about 500 pg / mL (e.g. about 501 pg / mL or above) in an (e.g., undiluted) blood sample. The concentration levels of BDG disclosed herein can be combined with any single concentration or any two or three concentrations of any other biomarkers, such as PCT, MxA and CRP.

[0100] In the test device, according to some embodiments of the invention, the fungal biomarker substance may comprise Galactomannan (GM). GM may be detected at a single concentration, or at two or more concentrations (e.g. a low and high concentration, or a low, medium and high concentration).

[0101] When the device is configured to detect GM at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) is visually determinable as bound to the immobilized binder via the fungal biomarker comprising GM, when a concentration of the fungal biomarker within the liquid sample corresponds to an optical density index, GDI, value of at least about 0.5 in an (e.g., undiluted) blood sample. The inventors have found that this concentration of GM biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to start to be reliably visually determined. The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result.

[0102] When the device is configured to detect GM at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the first detector reagent (e.g., coloured particles) bound to the immobilized binder via the fungal biomarker comprising GM saturates when a concentration of the fungal biomarker within the liquid sample corresponds to an optical density index, ODI, value of at least about 1 .5 in an (e.g., undiluted) blood sample. The inventors have found that this concentration of GM biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to create a maximally visually determinable accumulation of detector reagent (e.g., coloured particles). The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result.

[0103] When the device is configured so that GM is detected at two or more concentrations, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) is visually determinable as bound to the immobilized binder via the fungal biomarker comprising GM at a first line, when a concentration of the fungal biomarker within the liquid sample corresponds to an optical density index, ODI, value of about below 1 .5 in an (e.g., undiluted) blood sample and a second line when a concentration of the fungal biomarker within the liquid sample corresponds to an optical density index, ODI, value of about above 1 .5 in an (e.g., undiluted) blood sample. The accumulation of the detector reagent (e.g., coloured particles) may be visually determinable as bound to the immobilized binder via the fungal biomarker comprising GM at a first line, when a concentration of the fungal biomarker within the liquid sample corresponds to an optical density index, ODI, value of about 0.5-1 .4 in an (e.g., undiluted) blood sample and a second line when a concentration of the fungal biomarker within the liquid sample corresponds to an optical density index, ODI, value of about above 1 .5 in an (e.g., undiluted) blood sample. When the device is configured so that GM is detected at three or more concentrations, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) is visually determinable as bound to the immobilized binder via the fungal biomarker comprising GM at a first line, when a concentration of the fungal biomarker within the liquid sample corresponds to an optical density index, ODI, value of about below 1 .0 in an (e.g., undiluted) blood sample, at a second line when a concentration of the fungal biomarker within the liquid sample corresponds to an optical density index, ODI, of about 1.0-1 .4 in an (e.g., undiluted) blood sample, and a third line when a concentration of the fungal biomarker within the liquid sample corresponds to an optical density index, ODI, value of above about 1.5 in an (e.g., undiluted) blood sample. The concentration levels of GM disclosed herein can be combined with any single concentration or any two or three concentrations of any other biomarkers, such as PCT, MxA and CRP.

[0104] References herein to Optical Density, OD, may include a reference to optical Absorbance, as would be readily understood by the person of ordinary skill in the art.

[0105] The relative amount of light transmitted through a liquid sample may be defined as the ratio of the intensity of incident light, / 0, upon a sample, e.g., comprising GM, to the intensity of transmitted light, I, through the sample. This ratio may define a transmittance, T, of the analyte:

[0106] The absorbance, A, may be defined as:

[0107] The “optical density”, OD, may be defined as:

[0108] For absorbance measurements, the “optical density”, OD, may be so defined as a logarithmic measurement of the percent transmission (%T) in this way.

[0109] An optical density is preferably determined in respect of a particular wavelength of light corresponding to a peak, or maximum, absorbance value of the detector reagent within a spectral range of wavelengths of light of an absorbance spectrum of the detector reagent. For example, if gold nano-particles are used as detector reagent, then the spectral range of wavelengths may be from about 500nm to about 575nm, and the particular wavelength of light may be a value (e.g., one or 520nm, 530nm, 540nm, 560nm or 570nm) corresponding to the spectral position of an absorbance peak / maximum therein. The spectral position of an absorbance peak / maximum may depend upon the properties (e.g., size, material) of the detector reagent used. For example, a similar condition is discussed in relation to, for example, a detector reagent comprising gold nano-particles (cf. Fig.5 therein) in:

[0110] Manta et al.: "‘Optical Density Optimization of Malaria Pan Rapid Diagnostic Test Strips for Improved Test Zone Band Intensity’ Diagnostics 2020, 70(11), 880; https: / / doi.org / 10.3390 / diagnostics10110880

[0111] The same principle applies to other forms of detector reagent.

[0112] The absorbance of the second site (e.g., a test line area) in the presence of an analyte, e.g., GM, may be related to a concentration of the analyte according to a pre-set relation, known in the art as a calibration curve, which defines a relationship between analyte concentration and optical density of the second site (e.g., a test line area). The optical density is preferably determined under a condition that a relation between the concentration of GM and the absorbance value being measured is a substantially linear relation. For example, a similar condition is discussed (cf. Fig.1 therein) in:

[0113] Mercier et al.: “Galactomannan, a Surrogate Marker for Outcome in Invasive Aspergillosis: Finally Coming of Age”, Front. Microbiol., 04 April 2018, Volume 9 - 2018, https: / / doi.Org / 10.3389 / fmicb.2018.00661

[0114] Typically, Galactomannan tests are run using an immunoenzymatic sandwich microplate assay, such as for example, The Platelia™ Aspergillus EIA available from Bio-Rad. The test results from such assays are reported as GDI values and the GM clinical threshold values used throughout the literature, healthcare guidance and subsequently in this document are based on the GDI values established using this type of test. The assay works by first coating the wells of a microplate with monoclonal antibodies that are specific for Aspergillus galactomannan. The serum sample is then added to the wells and incubated. If Aspergillus galactomannan is present in the sample, it will bind to the antibodies on the microplate. Next, a conjugate reagent is added to the wells. This conjugate reagent is a mixture of monoclonal antibodies that are specific for Aspergillus galactomannan and that are linked to the enzyme peroxidase. The conjugate reagent binds to the Aspergillus galactomannan that is bound to the antibodies on the microplate. Finally, a substrate for peroxidase is added to the wells. This substrate reacts with the peroxidase in the conjugate reagent to produce a coloured product. The intensity of the colour is proportional to the amount of Aspergillus galactomannan that is present in the sample. An Optical Density (OD) value of the sample is then calculated by measuring the absorbance of the coloured product at 450 nm using a spectrophotometer.

[0115] The Optical Density, OD, value, as calculated by a spectrophotometer, is a measure of how much light is absorbed by a sample. The OD value is a measure of the concentration of a substance in a sample. The more concentrated a substance is, the more light it will absorb, and the higher the OD value will be.

[0116] An Optical Density value of a control or cut-off sample (provided by the manufacturer) is then also calculated by measuring the absorbance of this coloured product at 450 nm using the spectrophotometer. The Optical Density Index (ODI) is then calculated thus:

[0117] ODI = Optical Density of test sample / Optical Density of cut-off sample

[0118] Test samples with an ODI < 0.50 are considered to be negative for galactomannan antigen. Details can be found at in:

[0119] PLATELIA™ ASPERGILLUS EIA (96 TESTS 62796) - “THE PLATELIA™ ASPERGILLUS EIA IS AN IMMUNOENZYMATIC SANDWICH MICROPLATE ASSAY FOR THE DETECTION OF ASPERGILLUS GALACTOMANNAN ANTIGEN IN SERUM”, document code no. code: 881045, published by, and available from, Bio-Rad Laboratories Ltd, having an address at: The Junction, Station Road, Watford, Hertfordshire, WD17 1 ET, United Kingdom.

[0120] See: https: / / commerce.bio-rad.com / webroot / web / pdf / inserts / CDG / en / 62796_881045_EN.pdf

[0121] In the test device, according to some embodiments of the invention, the viral biomarker substance may comprise Myxovirus resistance protein A (MxA). MxA may be detected at a single concentration, or at two or more concentrations (e.g. a low and high concentration, or a low, medium and high concentration).

[0122] When the device is configured so that MxA is detected at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) is visually determinable as bound to the immobilized binder via the viral biomarker comprising MxA, when a concentration of the viral biomarker within the liquid sample corresponds to at least about 40ng / mL in an (e.g., undiluted) blood sample. The inventors have found that this concentration of MxA biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to start to be reliably visually determined. The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result.

[0123] When the device is configured so that MxA is detected at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) is visually determinable as bound to the immobilized binder via the viral biomarker comprising MxA, when a concentration of the viral biomarker within the liquid sample corresponds to at least about 80ng / mL in an (e.g., undiluted) blood sample. The inventors have found that this concentration of MxA biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to start to be reliably visually determined. The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result.

[0124] When the device is configured so that MxA is detected at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the further detector reagent (e.g., coloured particles) bound to the immobilized binder via the viral biomarker comprising MxA saturates when a concentration of the viral biomarker within the liquid sample corresponds to at least about 400ng / mL in an (e.g., undiluted) blood sample. The inventors have found that this concentration of MxA biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to create a maximally visually determinable accumulation of detector reagent (e.g., coloured particles). The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result.

[0125] When the device is configured so that MxA is detected at two or more concentrations, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the viral biomarker comprising MxA may saturate at a first test line when a concentration of the viral biomarker within the liquid sample corresponds to about 60 ng / mL or below in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the viral biomarker within the liquid sample corresponds to above 60 ng / mL or above (e.g. 61 ng / mL or above) in an (e.g., undiluted) blood sample. The accumulation of the detector reagent (e.g. coloured particles) bound to the immobilized binder via the viral biomarker comprising MxA may saturate at a first test line when a concentration of the viral biomarker within the liquid sample corresponds to about 60 ng / mL or above in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the viral biomarker within the liquid sample corresponds to about 80 ng / mL or above in an (e.g., undiluted) blood sample. The accumulation of the detector reagent (e.g. coloured particles) bound to the immobilized binder via the viral biomarker comprises MxA may saturate at a first test line when a concentration of the viral biomarker within the liquid sample corresponds to between about 40-60 ng / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the viral biomarker within the liquid sample corresponds to about 80 ng / mL or above in an (e.g., undiluted) blood sample. When the device is configured so that MxA is detected at three or more concentrations, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the viral biomarker comprising MxA may saturate at a first test line when a concentration of the viral biomarker within the liquid sample corresponds to below about 60 ng / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the viral biomarker within the liquid sample corresponds to between about 80-100 ng / mL in an (e.g., undiluted) blood sample and may saturate a third test line when a concentration of the viral biomarker within the liquid sample corresponds to above about 100 ng / mL (i.e. about 101 ng / mL or above) in an (e.g., undiluted) blood sample. The accumulation of the detector reagent (e.g. coloured particles) bound to the immobilized binder via the viral biomarker comprises MxA may saturate at a first test line when a concentration of the viral biomarker within the liquid sample corresponds to between about 40-60 ng / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the viral biomarker within the liquid sample corresponds to about 80-100 ng / mL in an (e.g., undiluted) blood sample and may saturate at a third test line when a concentration of the viral biomarker within the liquid sample corresponds to above about 100 ng / mL (i.e. about 101 ng / mL or above) in an (e.g., undiluted) blood sample. The concentration levels of MxA disclosed herein can be combined with any single concentration or any two or three concentrations of any other biomarkers, such as PCT, BDG, GM and CRP.

[0126] The first portion may comprise a conjugate comprising a binder for a non-specific inflammatory biomarker substance comprising C-Reactive Protein (CRP) coupled to a yet further respective detector reagent, and the second portion may comprise a binder immobilized therein which binds to CRP. Thus, for example, the test device may comprise a fourth conjugate for a non-specific inflammatory biomarker substance. Thus, the at least one further analyte may comprise a non-specific inflammatory biomarker. In the test device, according to some embodiments of the invention, the non-specific inflammatory biomarker substance may comprise C-Reactive Protein (CRP). CRP may be detected at a single concentration, or at two or more concentrations (e.g. a low and high concentration, or a low, medium and high concentration).

[0127] When the device is configured so that CRP is detected at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the yet further detector reagent (e.g., coloured particles) is visually determinable as bound to the immobilized binder via the non-specific inflammatory biomarker comprising CRP, when a concentration of the non-specific inflammatory biomarker within the liquid sample corresponds to at least about 10mg / L in an (e.g., undiluted) blood sample. The inventors have found that this concentration of CRP biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to start to be reliably visually determined. The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result.

[0128] When the device is configured so that CRP is detected at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the yet further detector reagent (e.g., coloured particles) bound to the immobilized binder via the non-specific inflammatory biomarker comprising CRP saturates when a concentration of the non-specific inflammatory biomarker within the liquid sample corresponds to at least about 100mg / L in an (e.g., undiluted) blood sample. The inventors have found that this concentration of CRP biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to create a maximally visually determinable accumulation of detector reagent (e.g., coloured particles). The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result.

[0129] When the device is configured so that CRP is detected at two or more concentrations, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the non-specific biomarker comprising CRP may saturate at a first test line when a concentration of the non-specific inflammatory biomarker within the liquid sample corresponds to below about 10.0 mg / dL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the non-specific inflammatory biomarker within the liquid sample corresponds to about 10.0 mg / dL or above in an (e.g., undiluted) blood sample. The accumulation of the detector reagent (e.g. coloured particles) bound to the immobilized binder via the non-specific inflammatory biomarker comprises CRP may saturate at a first test line when a concentration of the non-specific inflammatory biomarker within the liquid sample corresponds to between about 1 .0-9.9 mg / dL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the non-specific inflammatory biomarker within the liquid sample corresponds to about 10.0 mg / dL or above in an (e.g., undiluted) blood sample. When the device is configured so that CRP is detected at three or more concentrations, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the non-specific inflammatory biomarker comprising CRP may saturate at a first test line when a concentration of the non-specific inflammatory biomarker within the liquid sample corresponds to about 2.0 mg / dL or below in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the non-specific biomarker within the liquid sample corresponds to above about 2.0 mg / dL but below about 10.0 mg / dL in an (e.g., undiluted) blood sample and may saturate a third test line when a concentration of the non-specific inflammatory biomarker within the liquid sample corresponds to about 10.0 mg / dL or above in an (e.g., undiluted) blood sample. The accumulation of the detector reagent (e.g. coloured particles) bound to the immobilized binder via the non-specific inflammatory biomarker comprising CRP may saturate at a first test line when a concentration of the non-specific inflammatory biomarker within the liquid sample corresponds to between about 1 .0-2.0 mg / dL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the non-specific inflammatory biomarker within the liquid sample corresponds to about 2.1-9.9 mg / dL in an (e.g., undiluted) blood sample and may saturate at a third test line when a concentration of the non-specific inflammatory biomarker within the liquid sample corresponds to about 10.0 mg / dL or above in an (e.g., undiluted) blood sample. The concentration levels of CRP disclosed herein can be combined with any single concentration or any two or three concentrations of any other biomarkers, such as PCT, BDG, GM and MxA.

[0130] In the test device, according to some embodiments of the invention, the bacterial biomarker substance may comprise Procalcitonin (PCT). PCT may be detected at a single concentration, or at two or more concentrations (e.g. a low and high concentration, or a low, medium and high concentration). When the device is configured to detect PCT at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) is visually determinable as bound to the immobilized binder via the bacterial biomarker comprising PCT, when a concentration of the bacterial biomarker within the liquid sample corresponds to at least about 0.50 ng / mL in an (e.g., undiluted) blood sample. The inventors have found that this concentration of PCT biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to start to be reliably visually determined. The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result. This concentration of PCT may be particularly useful for Emergency, ICU or secondary care users of the device.

[0131] When the device is configured to detect PCT at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) is visually determinable as bound to the immobilized binder via the bacterial biomarker comprising PCT, when a concentration of the bacterial biomarker within the liquid sample corresponds to at least about 0.25 ng / mL in an (e.g., undiluted) blood sample. The inventors have found that this concentration of PCT biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to start to be reliably visually determined. The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result. This concentration of PCT may be particularly useful for community or primary care users of the device.

[0132] When the device is configured to detect PCT at a single concentration, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the at least one further detector reagent (e.g., coloured particles) bound to the immobilized binder via the bacterial biomarker comprising PCT saturates when a concentration of the bacterial biomarker within the liquid sample corresponds to at least about 5.0 ng / mL in an (e.g., undiluted) blood sample. The inventors have found that this concentration of PCT biomarker in a liquid sample comprising blood (e.g., undiluted) results in a sufficient accumulation of immobilized detector reagent (e.g., coloured particles) within the second portion (i.e., having become bound at the binder there) to create a maximally visually determinable accumulation of detector reagent (e.g., coloured particles). The concentration of the immobilized binder within the second portion may be chosen to be sufficient to achieve this result.

[0133] When the device is configured so that PCT is detected at two or more concentrations, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the bacterial biomarker comprising PCT may saturate at a first test line when a concentration of the bacterial biomarker within the liquid sample corresponds to below about 0.25 ng / mL or below about 0.50 ng / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the bacterial biomarker within the liquid sample corresponds to about 0.25 ng / mL or above, or about 0.50 ng / mL or above in an (e.g., undiluted) blood sample. The liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the bacterial biomarker comprising PCT may saturate at a first test line when a concentration of the bacterial biomarker within the liquid sample corresponds to below about 0.25 ng / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the bacterial biomarker within the liquid sample corresponds to about 0.25 ng / mL or above in an (e.g., undiluted) blood sample, e.g. when the test device is used in a community or primary care setting. The liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the bacterial biomarker comprising PCT may saturate at a first test line when a concentration of the bacterial biomarker within the liquid sample corresponds to below about 0.50 ng / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the bacterial biomarker within the liquid sample corresponds to about 0.50 ng / mL or above in an (e.g., undiluted) blood sample, e.g. when the test device is used in an ICU setting or secondary care setting. The liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the bacterial biomarker comprising PCT may saturate at a first test line when a concentration of the bacterial biomarker within the liquid sample corresponds to between about 0.10-0.24 ng / mL or about 0.10-0.49 ng / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the bacterial biomarker within the liquid sample corresponds to about 0.25 ng / mL or above, or about 0.50 ng / mL or above in an (e.g., undiluted) blood sample. The liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the bacterial biomarker comprising PCT may saturate at a first test line when a concentration of the bacterial biomarker within the liquid sample corresponds to between about 0.10-0.24 ng / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the bacterial biomarker within the liquid sample corresponds to about 0.25 ng / mL or above in an (e.g., undiluted) blood sample. The liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g., coloured particles) bound to the immobilized binder via the bacterial biomarker comprising PCT may saturate at a first test line when a concentration of the bacterial biomarker within the liquid sample corresponds to between about 0.10-0.49 ng / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the bacterial biomarker within the liquid sample corresponds to about 0.50 ng / mL or above in an (e.g., undiluted) blood sample. When the device is configured so that PCT is detected at three or more concentrations, the liquid sample may be (or comprise) a blood sample and the immobilized binder at the second portion may be configured such that an accumulation of the detector reagent (e.g. coloured particles) bound to the immobilized binder via the bacterial biomarker comprising PCT may saturate at a first test line when a concentration of the bacterial biomarker within the liquid sample corresponds to below about 0.25 ng / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the bacterial biomarker within the liquid sample corresponds to about 0.25 ng / mL or above and less than about 0.50 ng / mL in an (e.g., undiluted) blood sample, and may saturate at a third line when a concentration of the bacterial biomarker within the liquid sample corresponds to about 0.50 ng / mL or above in an (e g., undiluted) blood sample. The accumulation of the detector reagent (e.g. coloured particles) bound to the immobilized binder via the bacterial biomarker comprising PCT may saturate at a first test line when a concentration of the bacterial biomarker within the liquid sample corresponds to between about 0.10-0.24 ng / mL in an (e.g., undiluted) blood sample and may saturate at a second test line when a concentration of the bacterial biomarker within the liquid sample corresponds to about 0.25-0.49 ng / mL in an (e.g., undiluted) blood sample, and may saturate at a third line when a concentration of the bacterial biomarker within the liquid sample corresponds to about 0.50 ng / mL or above in an (e.g., undiluted) blood sample. The concentration levels of PCT disclosed herein can be combined with any single concentration or any two or three concentrations of any other biomarkers, such as BDG, GM, MxA and CRP.

[0134] The test device may comprise conjugates and respective immobilized binders for at least BDG (fungal) or GM (fungal), PCT (bacterial) and MxA (viral), wherein at least one of the biomarkers is detected at two or more concentrations. For example, the test device may comprise four respective test lines (of immobilized binders) in the second portion accordingly, wherein two test lines detect a single concentration of a biomarker respectively and two test lines detected two or more concentrations of a biomarker. For example, one test line detects a single concentration of BDG, one test line detects a single concentration of PCT, one test line detects a low concentration of MxA and one test line detects a high concentration of MxA. For example, one test line detects a single concentration of MxA, one test line detects a single concentration of PCT, one test line detects a low concentration of BDG and one test line detects a high concentration of BDG. For example, one test line detects a single concentration of MxA, one test line detects a single concentration of BDG, one test line detects a low concentration of PCT and one test line detects a high concentration of PCT. For example, the test device may comprise five respective test lines (of immobilized binders) in the second portion accordingly, wherein one test lines detects a single concentration of a biomarker and four test lines detects two or more concentrations of a two biomarkers. For example, one test line detects a single concentration of BDG, one test line detects a low concentration of PCT, one test line detects a high concentration of PCT, one test line detects a low concentration of MxA and one test line detects a high concentration of MxA. For example, one test line detects a single concentration of MxA, one test line detects a low concentration of PCT, one test line detects a high concentration of PCT, one test line detects a low concentration of BDG and one test line detects a high concentration of BDG. For example, one test line detects a single concentration of PCT, one test line detects a low concentration of MxA, one test line detects a high concentration of MxA, one test line detects a low concentration of BDG and one test line detects a high concentration of BDG. For example, the test device may comprise six respective test lines (of immobilized binders) in the second portion accordingly, wherein one test line detects a high concentration of MxA, one test line detects a low concentration of MxA, one test line detects a high concentration of PCT, one test line detects a low concentration of PCT, one test line detects a low concentration of BDG and one test line detects a high concentration of BDG. The device may comprise a further test line for each analyte if the device is configured to also detect a medium concentration of one or more of the analytes. The test device may comprise conjugates and respective immobilized binders for at least BDG (fungal) or GM (fungal), PCT (bacterial) and MxA (viral) and CRP (non-specific inflammatory biomarker). CRP may be detected at a single concentration or at two or more concentrations. The test device may comprise five respective test lines (of immobilized binders) in the second portion accordingly, with additional test lines (binding the non-specific inflammatory biomarker comprising CRP and its respectively co-attached conjugate) being useful to confirm that there is some inflammation going on. The device further comprises one additional test line if CRP is detected at a single concentration. The device further comprises two additional test lines if CRP is detected at a first and second (e.g. low and high) concentration. The device further comprises three test lines if CRP is detected at a first, second and third (e.g. low, medium and high) concentration.

[0135] Capture reagents (i.e., said binders) may be applied in a suitable medium to provide consistent application and functionality once bound. For example, a 0.1 Molar phosphate buffer at pH 7.4 may be used. For example, a compound of sodium phosphate dibasic heptahydrate at 0.07541 M may be used, or a compound of sodium phosphate monobasic monohydrate at 0.02459 M may be used. Other buffering systems could be utilised for example Tris based buffers and the inclusion of other specific agents can be utilised if needed, for example but not limited to bovine serum albumin, sugars or other agents to control non-specific binding or to improve reagent stability.

[0136] Preferably, only when the concentration of the biomarker in the sample exceeds its pre-set threshold value (e.g. single concentration level listed above for each biomarker, or low and high (and optionally medium) concentration level listed above for each biomarker) will a positive test line be visibly displayed (i.e. it ignores the corresponding “normal” levels of each biomarker in the blood sample). This gives the user a “go / no-go” type result rather than a line intensity result. The ability to not require a separate test reader has certain advantages in the field of use such as cost reduction, zero calibration / training requirements and improved accessibility at the point of need. When the device is configured to detect a low and high (and optionally medium) concentration level for a given biomarker, the clinician can immediately determine the severity of the infection and / or inflammation and determine an appropriate treatment.

[0137] Above the threshold (e.g. concentration) levels the intensity of the test line preferably increases in proportion to the concentration of the biomarker in the sample, so it can also be used with a test reader should the user require some form of semi-quantitative understanding of the severity of infection (if present) for test device configurations employing single concentration test lines i.e. where the test lines are not configured to indicate multiple defined concentration values of the biomarker(s). But in the initial triage of sepsis diagnosis in more advanced secondary care settings this is not so critical as further blood tests, cultures etc. will still be available and may be used to refine and monitor treatment if a test reader is not present. The test device may in any case be used without a test reader and still allows a user to focus on the immediate initial diagnosis of sepsis and what type (bacterial, fungal or viral) which is key to saving lives and improving outcomes. In a second aspect, the invention may provide a test device as disclosed herein wherein the permeable material is planar and the first site is laterally spaced from the second site along the planar permeable material such that the test device provides a lateral flow test device. For example, a lateral flow test device may be provided for detecting elevated levels of CRP, PCT and BDG, wherein at least one of the biomarkers is detected at two or more concentration levels to aid the early determination of the trigger of the inflammatory response. For example, a lateral flow test device may be provided for detecting elevated levels of PCT, MxA and BDG and / or GM and optionally CRP, wherein at least one of the biomarkers is detected at two or more concentration levels, to aid the early determination of the trigger of the inflammatory response.

[0138] In a further aspect, the invention may provide a diagnostic device forthe early identification of microbial infection types, in biological samples. The invention may relate to diagnostic devices which aid rapid diagnosis of infection and disease. Particularly, although not exclusively, the invention may relate to the early identification of the type of infection (bacterial, fungal or viral) triggering a dysregulated immune response in suspected sepsis cases. In a general aspect, the invention may provide a diagnostic device for the detection of both bacterial and fungal infections in humans.

[0139] In a further general aspect, the invention may provide a single diagnostic device for detection of elevated levels (or clinically significant levels, e.g. high concentration levels) of Procalcitonin (PCT), C-reactive protein (CRP) and (1-3)-p-D-glucan (BDG) in a biological sample, including (but not limited to) blood, sweat, sputum, mucus, urine or saliva. A biological sample may also include serum or plasma.

[0140] In another, further general aspect the invention may provide the ability to rapidly identify possible sepsis cases in humans at the point of care.

[0141] In another, further general aspect the invention may provide the ability to rapidly differentiate infective critical illness e.g. sepsis, from non-infective critical illness e.g. Systemic Inflammatory Response Syndrome (SIRS) in humans at the point of care.

[0142] Furthermore, the device may be configured to identify an elevated general inflammatory response (as indicated by elevated levels of CRP, e.g. high concentration levels) as well as distinguish between bacterial sepsis (as indicated by elevated levels of PCT) and fungal sepsis (as indicated by elevated levels of BDG) and thereby also giving guidance to the possibility of viral infection. Furthermore, the device may be configured to identify a general inflammatory response and the possibility of a viral infection, as indicated by elevated levels of CRP (along with the patient’s recent medical history) as well as detecting and distinguishing bacterial infections (as indicated by elevated levels of PCT) and fungal infections (as indicated by elevated levels of BDG).

[0143] In another general aspect, the invention may provide the detection of elevated levels of PCT, BDG and CRP using a lateral flow device (LFD). Such device may be either a sandwich immunoassay or competitive assay, with both methods being known to those skilled in the art. The device may implement inhibition assay such as is known to those skilled in the art. The test being either in a dipstick or cassettetype format (for example, enclosed in a plastic housing).

[0144] In another general aspect, the invention may provide a lateral flow device for the combined detection of Procalcitonin (PCT) and (1-3)-p-d-glucan (BDG) and optionally C-Reactive Protein (CRP) in biological samples, wherein at least one of PCT and BDG is detected at two or more concentration levels (e.g. low / high concentrations or low / medium / high concentration).

[0145] In another general aspect, the invention may provide a lateral flow device for the combined detection of Procalcitonin (PCT) and Myxovirus resistance protein (MxA) and optionally C-Reactive Protein (CRP), in biological samples, wherein at least one of PCT and MxA is detected at two or more concentration levels(e.g. low / high concentrations or low / medium / high concentration)

[0146] The detection of the target biomarkers (e.g. PCT, CRP and BDG) may be conducted using the same lateral flow device, i.e. a multiplex lateral flow device. Said detection of biomarkers at a single concentration level or at two or more concentration levels (e.g. low / high concentrations or low / medium / high concentration) may be performed on either a single test channel or on two or more separate test channels within the same device, depending on the materials and reagents chosen and their relative sensitivities for each biomarker. Said test channels may share or have their own sample entry port for receiving the biological sample under investigation. For example, a low concentration of BDG may be detected on one test channel and a high concentration of BDG may be detected on a separate test channel. Alternatively, the two or more concentrations of BDG are detected on the same test channel. In some configurations, all of the biomarkers being detected within the sample are detected (at a single or at two or more concentrations) within the same channel. In other configurations, each of the individual biomarkers being detected are separated into separate test channels.

[0147] The test sample may be either unprepared (as extracted from its source) or prepared (mixed with reagents to enhance the stability and / or the performance / detectability of the sample during the test). Examples of sample and test enhancing reagents may include, but are not limited to, pH mediating buffers, diluents, viscosity control fluids, enzymes, antibodies, and agglutination agents.

[0148] Further examples of sample and test enhancing reagents may include selective lysing agents, such as leukocyte membrane lysing agents.

[0149] Sample and test enhancing reagents may be employed either prior to sample application or in situ once the sample has been applied to the device.

[0150] In another general aspect, the invention may provide the use of antibodies which have been conjugated to nanoparticles for the selective binding and detection of each biomarker (e.g. CRP, PCT and BDG). Said nanoparticles providing the visual indication of the presence of each biomarker when captured at its specific test line. Examples of suitable nanoparticles include, but are not limited to, gold, carbon or latex. The invention may provide the use of said binders (also known as binding agents) on a form comprising (but not limited to): antibodies, antibody fragments, antigens, enzymes, aptamers or affimers. The binders are preferably conjugated to a suitable detection reagent or molecular responder system such as (but not limited to): nanoparticles; nanoshells; enzyme linked systems. These may provide a selective binding and detection of each individual biomarker (e.g., CRP, PCT, BDG etc.). The detection reagent or molecular responder system may provide the visual indication of the presence of each biomarker when captured in the second portion of the test device e.g., at its specific test line. Examples of suitable nanoparticle detection reagents include, but are not limited to: gold, carbon or latex.

[0151] The test device may provide the ability to establish an individual minimum response or threshold concentration value for each biomarker (i.e. as a single test line for a single concentration). By doing this, the test may confirm the presence of a specific biomarker (e.g., by displaying a visible line at the relevant test line) if it is present in the sample above the threshold concentration value set for that particular biomarker on the test device. In this way, the test may be configured to ignore (or be visually unresponsive to) low level or clinically normal concentrations of biomarkers and can be tuned to only display clinically significant levels as required for each clinical application and / or sample type. As disclosed herein, the test is configured to test the presence of at least one of the viral, fungal or bacterial biomarkers at two or more concentration levels (e.g. low / high concentrations or low / medium / high concentration). The test device may detect two concentrations (e.g. low / high) for one biomarker and three concentrations (e.g. low / medium / high) for another biomarker.

[0152] In a further aspect, the invention may provide that the intensity of each test line is proportional to the concentration of each biomarker in the sample, therefore the test is not only qualitative but also quantitative (if required). The quantitative accuracy of the test may be enhanced by use of a controlled sample volume, magnetic or fluorescent labelled antibody conjugates along with a suitable companion lateral flow device reader, mobile device app, or camera, or meter and applicable transformative data algorithms.

[0153] Potential designs and materials that may be used to create a suitable multiplexed lateral flow device such as, for example, a sandwich type assay, will be obvious to those skilled in the art. Generally speaking, such a device may comprise a number of distinct zones, the first of which may be a sample pad (or pads) which is / are designed to accept the test sample, filter any unwanted sample components (for example, in the case of a whole blood sample, removing red blood cells) and to wick the sample towards an adjoined conjugate pad.

[0154] The conjugate pad (or pads) typically may contain the key test reagents, such as, but not limited to, biomarker specific nanoparticle-antibody conjugates and the control line specific conjugates. It is here that the biomarkers (if present in the sample) bind to their specific nanoparticle-antibody conjugate to form a biomarker conjugate complex.

[0155] The conjugate pad (or pads) may contain the key test reagents, such as (but not limited to): biomarker specific nanoparticle-antibody conjugates (or other specific binding agent and detection system) and any control line specific conjugates (if required). Biomarkers (if present in the sample) may bind to their specific nanoparticle-antibody conjugates to form biomarker-conjugate complexes.

[0156] A test zone may be provided and may adjoin the conjugate pad which may comprise a suitable wicking media to draw the sample into this zone, such as, but not limited to, a nitrocellulose membrane.

[0157] Distinct test lines (one or more test lines) for each biomarker may be defined in this region. Desirably, only a single set of antibodies (or other biomarker specific capture reagent or feature) are immobilised at each test line, thereby allowing for selective capture of the corresponding biomarker conjugate complex at the relevant designated test line. For example, at the first test line, optionally only the CRP conjugate complex is captured. At a second test line, optionally only the PCT conjugate complex is captured. At the third test line, optionally only the BDG conjugate complex is captured. At a control line, optionally only the control line conjugate is captured. In some examples, excess biomarker conjugates may be captured to create a visible control line.

[0158] The control line may be a separate line beyond the test lines and may provide a visual indication that the test has been conducted successfully (or not).

[0159] An absorbent pad may be included at the end of the device to ensure the sample is drawn to the very end of the device and to capture any excess sample once past the test zone.

[0160] The detection of at least one of the viral, fungal or bacterial biomarkers at two or more concentration levels (e.g. low / high concentrations or low / medium / high concentration) is achieved by configuring the test lines within the second portion to display a visible line once the first and second (and optionally third) concentration levels of a biomarker are present in the sample. For example, the first test line may be configured to display a visible test line at the lower biomarker concentration. The second test line may be configured to display a visible test line at the higher biomarker concentration. If a third test line is present, then the third test line may be configured to display a visible test line at the highest biomarker concentration and the second test line configured to display a visible test line at a biomarker concentration that is between the first test line and third test line biomarker concentrations. In some configurations, the test lines are ordered such that the first test line (e.g. low concentration test line) is closest to the sample pad and the second test line (e.g. high concentration test line) is furthest away from the sample pad. In some configurations, the test lines are ordered such that the second test line (e.g. high concentration test line) is closest to the sample pad and the first test line (e.g. low concentration test line) is furthest away from the sample pad. If a medium concentration test line is present, then this test line may fall between the low and high test lines. In this way, the test lines may become visible in order of increasing sample biomarker concentration values running either from closest to the sample pad to furthest away (e.g. an upwards visible ladder scale), or from furthest away from the sample pad to the closest (e.g. a downwards visible ladder scale). For example, in some configurations a series of test lines may be configured on the second portion (e.g. test zone) of the device whereby the concentration of the immobilised binders at each test line are set to allow a controlled amount (concentration) of the biomarker and / or biomarker conjugate complex to progress from one test line to the next test line above a predetermined biomarker concentration level, e.g. from the first test line towards the second and / or third test line(s), which are co-configured to capture any excess biomarker in the sample not captured by the preceding test line(s), to a desired concentration level, thereby visibly signalling the presence of biomarker to the desired test sample concentration level for that particular test line. For example, in other configurations, a series of test lines may be created on the second portion (e.g. test zone) of the device with increasing concentration of the immobilised binders at each test line or immobilised binders with increasing affinity for the biomarker conjugate complex at each test line. The test lines only display a visible test line once the test line specific concentration is met by the relevant concentration of the biomarker in the sample. These and other such methods and configurations that may be utilised to produce the desire effect are known to those skilled in the art.

[0161] The test line may be a continuous or a non-continuous test line formed of immobilised binder. For example, the test line may be a discrete arrangement of immobilised binder in the form of a collection of dots or microdots of various shapes and / or sizes.

[0162] The test line may be orientated and arranged in any direction e g. parallel to the sample flow, or perpendicular to the sample flow, or diagonally. The test line may be of any shape, such as, for example a lineartest line. Alternatively, the test line may be a circular, e.g. have a circular pattern. Alternatively, the test line may be a combination of different shapes, including, but not limited to a combination of linear and circular test lines.

[0163] The invention also provides methods of detecting sepsis in a sample obtained from a subject using the test device described herein. The methods of detecting sepsis may comprises detecting a fungal biomarker and a viral biomarker (e.g. BDG and MxA), wherein at least one or both of the biomarkers are detected at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high). The methods of detecting sepsis may comprises detecting a fungal biomarker and a bacterial biomarker (e.g. BDG and PCT), wherein at least one or both of the biomarkers are detected at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high). The methods of detecting sepsis may comprise detecting a viral biomarker and a bacterial biomarker (e.g. PCT and MxA) wherein at least one or both of the biomarkers are detected at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high). The methods of detecting sepsis disclosed herein may further comprise detecting a non-specific inflammatory marker in a sample obtained from a subject using the test device described herein. The non-specific inflammatory marker may be CRP and may be detected at a single concentration, or at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high). For example, the methods of detecting sepsis comprise detecting BDG, PCT and MxA, wherein at least one or more of the biomarkers are detected at two or more different concentrations (e.g. two concentrations such as low / high, e g. three concentrations, such as low / medium / high), optionally in combination with CRP, which may be detected at a single concentration, or at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high). In another example, the methods of detecting sepsis comprises detecting GM, PCT and MxA wherein at least one or more of the biomarkers are detected at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high), optionally in combination with CRP, which may be detected at a single concentration, or at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high).

[0164] The invention also provides methods of selecting a treatment for a subject, for example, a subject suspected of having sepsis, using the test device described herein. The methods of selecting a treatment for a subject may comprises detecting a fungal biomarker and a viral biomarker (e.g. BDG and MxA), wherein at least one or both of the biomarkers are detected at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high). The methods of selecting a treatment for a subject may comprises detecting a fungal biomarker and a bacterial biomarker (e.g. BDG and PCT), wherein at least one or both of the biomarkers are detected at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high). The methods of selecting a treatment for a subject may comprise detecting a viral biomarker and a bacterial biomarker (e.g. PCT and MxA) wherein at least one or both of the biomarkers are detected at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high). The methods of selecting a treatment for a subject disclosed herein may further comprise detecting a non-specific inflammatory marker in a sample obtained from a subject using the test device described herein. The non-specific inflammatory marker may be CRP and may be detected at a single concentration, or at two or more different concentrations (e.g. two concentrations such as low / high, e g. three concentrations, such as low / medium / high). For example, the methods of selecting a treatment for a subject comprise detecting BDG, PCT and MxA, wherein at least one or both of the biomarkers are detected at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high), optionally in combination with CRP, which may be detected at a single concentration, or at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high). In another example, the methods of selecting a treatment for a subject comprises detecting GM, PCT and MxA wherein at least one or more of the biomarkers are detected at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high), optionally in combination with CRP, may be detected at a single concentration, or at two or more different concentrations (e.g. two concentrations such as low / high, e.g. three concentrations, such as low / medium / high). The invention also provides methods of selecting a treatment for a subject, for example, a subject suspected of having sepsis, using the test device described herein.

[0165] The invention includes the combination of the aspects and preferred features described except where such a combination is clearly impermissible or expressly avoided.

[0166] Representative clauses

[0167] The invention will now be defined by the following clauses, which can all be combined with each other except where such a combination is clearly impermissible or expressly avoided.

[0168] 1 . A test device for detecting the presence of biomarkers in a liquid sample, the device comprising: a permeable material defining: a first portion providing a first site for application of the liquid sample and for conjugates movably supported therein wherein each conjugate comprises a binder for a respective biomarker coupled to a respective detector reagent; and, a second portion configured relative to the first portion so as to permit capillary flow communication therebetween, and said second portion providing a second site spaced from the first site for visually determining the presence of the respective detector reagent, and comprising binders immobilized therein each of which binds to a respective said biomarker; wherein the first portion comprises for a first said biomarker, a first said moveably supported conjugate comprising a first binder for a fungal, viral or bacterial biomarker substance coupled to a first respective detector reagent and, for at least one further said biomarker, at least one further said moveably supported conjugate comprising at least one further binder coupled to at least one further detector reagent, wherein the at least one further biomarker comprises a respective biomarker substance selected from a fungal, viral or bacterial biomarker substance, wherein the first biomarker and the at least one further biomarker are different biomarkers, and wherein the second site is configured to visually determine the presence of at least one of the respective detector reagents at two or more different concentrations of at least one of the biomarkers.

[0169] 2. The test device according to clause 1 , wherein the first portion comprises a first binder for a fungal biomarker substance and at least one further binder for a viral biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal biomarker substance at two or more different concentrations of the fungal biomarker substance.

[0170] 3. The test device according to clause 1 , wherein the first portion comprises a first binder for a fungal biomarker substance and at least one further binder for a viral biomarker substance, and wherein the second site is configured to visually determine the presence of the viral biomarker substance at two or more different concentrations of the viral biomarker substance.

[0171] 4. The test device according to clause 1 , wherein the first portion comprises a first binder for a fungal biomarker substance and at least one further binder for a viral biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal and viral biomarker substance at two or more different concentrations of the fungal or viral biomarker substance respectively.

[0172] 5. The test device according to clause 1 , wherein the first portion comprises a first binder for fungal biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal biomarker substance at two or more different concentrations of the fungal biomarker substance.

[0173] 6. The test device according to clause 1 , wherein the first portion comprises a first binder for a fungal biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the bacterial biomarker substance at two or more different concentrations of the bacterial biomarker substance. 7. The test device according to clause 1 , wherein the first portion comprises a first binder for a fungal biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal and bacterial biomarker substance at two or more different concentrations of the fungal and bacterial biomarker substance respectively.

[0174] 8. The test device according to clause 1 , wherein the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder binds for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal biomarker substance at two or more different concentrations of the fungal biomarker substance.

[0175] 9. The test device according to clause 1 , wherein the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the bacterial biomarker substance at two or more different concentrations of the bacterial biomarker substance.

[0176] 10. The test device according to clause 1 , wherein the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the viral biomarker substance at two or more different concentrations of the viral biomarker substance.

[0177] 11 . The test device according to clause 1 , wherein the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal and bacterial biomarker substance at two or more different concentrations of the fungal and bacterial biomarker substance respectively.

[0178] 12. The test device according to clause 1 , wherein the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal and viral biomarker substance at two or more different concentrations of the fungal and viral biomarker substance respectively.

[0179] 13. The test device according to clause 1 , wherein the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the bacterial and viral biomarker substance at two or more different concentrations of the bacterial and viral biomarker substance respectively. 14. The test device according to clause 1 , wherein the device detects the presence of three biomarkers, wherein the first portion comprises a binder for a fungal biomarker substance, a second binder binds for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal, bacterial and viral biomarker substance at two or more different concentrations of the fungal, bacterial and viral biomarker substance respectively.

[0180] 15. The test device according to clause 1 , wherein the first portion comprising a first binder for a viral biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the viral biomarker substance at two or more different concentrations of the viral biomarker substance.

[0181] 16. The test device according to clause 1 , wherein the first portion comprising a first binder for a viral biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the bacterial biomarker substance at two or more different concentrations of the bacterial biomarker substance.

[0182] 16a. The test device according to clause 1 , wherein the first portion comprising a first binder for a viral biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the bacterial biomarker substance at two or more different concentrations of the bacterial biomarker substance and visually determine the presence of the viral bacterial biomarker substance at two or more different concentrations of the viral biomarker substance.

[0183] 17. The test device according to any one of clauses 1-16a, wherein the first portion further comprises a binder for a non-specific inflammatory biomarker substance and wherein the second site is configured to visually determine the presence of the non-specific inflammatory biomarker at one concentration or at two or more different concentrations of the non-specific inflammatory biomarker.

[0184] 18. The test device according to any one of clauses 1-17, wherein the fungal biomarker substance comprises Beta-D-glucan (BDG).

[0185] 19. The test device according to any one of clauses 1-17, wherein the fungal biomarker substance comprises Galactomannan (GM).

[0186] 20. The test device according to any one of clauses 1-19, wherein the viral biomarker substance comprises Myxovirus resistance protein A (MxA).

[0187] 21. The test device according to any one of clauses 1-20, wherein the bacterial biomarker substance comprises Procalcitonin (PCT).

[0188] 22. The test device according to any one of clauses 1-21 , wherein the second site comprises two or more test lines comprising binders immobilised therein which bind to at least one respective biomarker, wherein each test line provides a visual determination of the presence of at least one biomarker at two or more different concentrations of said biomarker. 23. The test device according to clause 22, wherein the second site comprises two test lines for visually determining the presence of at least one biomarker at two or more different concentrations of the biomarker, wherein the first test line provides a visual determination of the presence of said at least one biomarker at a first concentration and the second test line provides a visual determination of the presence of said at least one biomarker at a second concentration.

[0189] 24. The test device according to clause 23, wherein the second site comprises two test lines for visually determining the presence of at least one biomarker at two or more different concentrations, wherein the first test line provides a visual determination of the presence of said at least one biomarker at a low concentration and the second test line provides a visual determination of the presence of said at least one biomarker at a high concentration.

[0190] 25. The test device according to any one of clauses 22-24, wherein the first portion comprises a first binder for a fungal biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal biomarker in the sample, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample.

[0191] 26. The test device according to any one of clauses 22-24, wherein the first portion comprises a first binder for a fungal biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal biomarker in the sample, wherein the fungal biomarker substance comprises GM and wherein the first test line provides a visual determination of the presence of GM corresponding to an optical density index, ODI, value of at least about 0.5-1 .4 in a blood sample and the second test line provides a visual determination of GM corresponding to an optical density index, ODI, value of at least above about 1 .5 in a blood sample.

[0192] 27. The test device according to any one of clauses 22-24, wherein the first portion comprises a first binder for a viral biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the viral biomarker substance in the sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample.

[0193] 28. The test device according to any one of clauses 22-24, wherein the first portion comprises a first binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL or below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above or about 0.50 ng / mL or above in a blood sample.

[0194] 29. The test device according to any one of clauses 22-24, wherein the first portion comprises a first binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above in a blood sample.

[0195] 30. The test device according to any one of clauses 22-24, wherein the first portion comprises a first binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.50 ng / mL or above in a blood sample.

[0196] 30a. The test device according to any one of clauses 25-30, wherein the first portion further comprises a binder for a non-specific inflammatory biomarker substance, and wherein the second site further comprises two or more test lines for visually determining the presence of the non-specific inflammatory biomarker substance in the sample, wherein the non-specific inflammatory biomarker substance comprises CRP and wherein the first test line provides a visual determination of the presence of CRP corresponding to a concentration of below about 9.9 mg / dL in a blood sample and the second test line provides a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

[0197] 31 . The test device according to clause 22, wherein the second site comprises three test lines comprising binders immobilised therein which bind to at least one respective biomarker, wherein each test line provides a visual determination of the presence of said biomarker at three or more different concentrations, wherein the first test line provides a visual determination of the presence of said at least one biomarker at a first concentration, the second test line provides a visual determination of the presence of said at least one biomarker at a second concentration, and the third test line provides a visual determination of the presence of said at least one biomarker at a third concentration.

[0198] 32. The test device according to clause 31 , wherein the second site comprises three test lines for visually determining the presence of at least one biomarker at three or more different concentrations, wherein the first test line provides a visual determination of the presence of said at least one biomarker at a low concentration, the second test line provides a visual determination of the presence of said at least one biomarker at a medium concentration, and the third test line provides a visual determination of the presence of said at least one biomarker at a high concentration. 33. The test device according to any one of clauses 31 or 32, wherein the first portion comprises a first binder for a fungal biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the fungal biomarker substance in the sample, wherein the fungal biomarker comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample, the second test line provides a visual determination of BDG corresponding to a concentration of about 81-500 pg / mL in a blood sample and the third test line provides a visual determination of BDG corresponding to a concentration of above about 500 pg / mL in a blood sample.

[0199] 34. The test device according to any one of clauses 31 or 32, wherein the first portion comprises a first binder for a fungal biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the fungal biomarker substance in the sample, wherein the fungal biomarker comprises GM and wherein the first test line provides a visual determination of the presence of GM corresponding to an optical density index, ODI, value corresponding to below about 1 .0 in a blood sample, the second test line provides a visual determination of GM corresponding to an optical density index, ODI, value of about 1 .0-1 .4 in a blood sample, and the third test line provides a visual determination of GM corresponding to an optical density index, ODI value of above about 1 .5 in a blood sample.

[0200] 35. The test device according to any one of clauses 31 or 32, wherein the first portion comprises a first binder for a viral biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the viral biomarker substance in the sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample, the second test line provides a visual determination of the presence of MxA corresponding to a concentration of about 61-100 ng / mL in a blood sample and the third test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 100 ng / mL in a blood sample.

[0201] 36. The test device according to any one of clauses 31 or 32, wherein the first portion comprises a first binder for a bacterial biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance comprises PCT, wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample, the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25-0.49 ng / mL in a blood sample and the third test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.5 ng / mL or above in a blood sample.

[0202] 36a. The test device according to any one of clauses 33-36, wherein the first portion further comprises a binder for a non-specific inflammatory biomarker substance, and wherein the second site further comprises three or more test lines for visually determining the presence of the non-specific inflammatory biomarker substance in the sample, wherein the non-specific inflammatory biomarker substance comprises CRP and wherein the first test line provides a visual determination of the presence of CRP corresponding to a concentration of below about 2.0 mg / dL in a blood sample, the second test line provides a visual determination of the presence of CRP corresponding to a concentration of about 2.1-9.9 mg / dL in a blood sample and the third test line provides a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

[0203] 37. The test device according to clause 22, wherein the second site comprises two or more test lines comprising binders immobilised therein for visually determining the presence of at least two biomarkers at two or more different concentrations for each biomarker, wherein each test line provides a visual determination of the presence of each biomarker at two or more different concentrations of each biomarker.

[0204] 38. The test device according to clause 37, wherein the second site comprises two or more test lines comprising binders immobilised therein which bind to at least two biomarkers for visually determining the presence of at least two biomarkers at two or more different concentrations for each biomarker, wherein the first test line for each biomarker provides a visual determination of the presence of the biomarker at first concentration for the biomarker and the second test line for each biomarker provides a visual determination of the presence of the biomarker at a second concentration for the biomarker.

[0205] 39. The test device according to any one of clauses 37 or 38, wherein the first portion comprises a first binder for a fungal biomarker substance and at least a further binder for a viral biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal and viral biomarker in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, and wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample.

[0206] 40. The test device according to any one of clauses 37 or 38, wherein the first portion comprises a first binder for a fungal biomarker substance and at least a further binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal and bacterial biomarker in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, and wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL or below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above or about 0.50 ng / mL or above in a blood sample. 41 . The test device according to any one of clauses 37 or 38, wherein the first portion comprises a first binder for a fungal biomarker substance and at least a further binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal and bacterial biomarker in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, and wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above in a blood sample.

[0207] 42. The test device according to any one of clauses 37 or 38, wherein the first portion comprises a first binder for a fungal biomarker substance and at least a further binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal and bacterial biomarker in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, and wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.50 ng / mL or above in a blood sample.

[0208] 43. The test device according to any one of clauses 37 or 38, wherein the first portion comprises a first binder for a viral biomarker substance and at least a further binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the bacterial and viral biomarker in the sample each at two or more different concentrations, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample and wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL or below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above or about 0.50 ng / mL or above in a blood sample.

[0209] 44. The test device according to any one of clauses 37 or 38, wherein the first portion comprises a first binder for a viral biomarker substance and at least a further binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the bacterial and viral biomarker in the sample each at two or more different concentrations, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample and wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above in a blood sample.

[0210] 45. The test device according to any one of clauses 37 or 38, wherein the first portion comprises a first binder for a viral biomarker substance and at least a further binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the bacterial and viral biomarker in the sample each at two or more different concentrations, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample and wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.50 ng / mL or above in a blood sample.

[0211] 46. The test device according to clause 22, wherein the second site comprises three test lines comprising binders immobilised therein which bind to at least two biomarkers for visually determining the presence of at least two biomarkers at three or more different concentrations for each biomarker, wherein each test line provides a visual determination of the presence of each biomarker at three or more different concentrations of each biomarker.

[0212] 47. The test device according to clause 46, wherein the second site comprises three test lines comprising binders immobilised therein which bind to at least two biomarkers for visually determining the presence of at least two biomarkers at three or more different concentrations for each biomarker, the first test line provides a visual determination of the presence of each biomarker at a first concentration for each biomarker, the second test line provides a visual determination of the presence of each biomarker at a second concentration for each biomarker and the third test line provides a visual determination of the presence of each biomarker at a third concentration for each biomarker.

[0213] 48. The test device according to clause 47, wherein the second site comprises three test lines comprising binders immobilised therein which bind to at least two biomarkers for visually determining the presence of at least two biomarkers at three or more different concentrations for each biomarker, the first test line provides a visual determination of the presence of each biomarker at a low concentration for each biomarker, the second test line provides a visual determination of the presence of each biomarker at a medium concentration for each biomarker and the third test line provides a visual determination of the presence of each biomarker at a high concentration for each biomarker.

[0214] 49. The test device according to any one of clauses 46-48, wherein the first portion comprises a first binder for a fungal biomarker substance and a viral biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the fungal and viral biomarker substance in the sample each at three different concentrations, wherein the fungal biomarker comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample, the second test line provides a visual determination of BDG corresponding to a concentration of about 81-500 pg / mL in a blood sample and the third test line provides a visual determination of BDG corresponding to a concentration of above about 500 pg / mL in a blood sample, and wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample, the second test line provides a visual determination of the presence of MxA corresponding to a concentration of about 61-100 ng / mL in a blood sample and the third test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 100 ng / mL in a blood sample.

[0215] 50. The test device according to any one of clauses 46-48, wherein the first portion comprises a first binder for a fungal biomarker substance and a bacterial biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the fungal and bacterial biomarker substance in the sample each at three different concentrations, wherein the fungal biomarker comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample, the second test line provides a visual determination of BDG corresponding to a concentration of about 81-500 pg / mL in a blood sample and the third test line provides a visual determination of BDG corresponding to a concentration of above about 500 pg / mL in a blood sample, and wherein the bacterial biomarker substance comprises PCT, wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample, the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25- 0.49 ng / mL in a blood sample and the third test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.5 ng / mL or above in a blood sample.

[0216] 51 . The test device according to any one of clauses 46-48, wherein the first portion comprises a first binder for a viral biomarker substance and a bacterial biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the viral and bacterial biomarker substance in the sample each at three different concentrations, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample, the second test line provides a visual determination of the presence of MxA corresponding to a concentration of about 61- 100 ng / mL in a blood sample and the third test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 100 ng / mL in a blood sample, and wherein the bacterial biomarker substance comprises PCT, wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample, the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25-0.49 ng / mL in a blood sample and the third test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.5 ng / mL or above in a blood sample.

[0217] 52. The test device according to clause 22, wherein the second site comprises two or more test lines comprising binders immobilised therein for visually determining the presence of at least three biomarkers at two or more different concentrations for each biomarker, wherein each test line provides a visual determination of the presence of each biomarker at two or more different concentrations of each biomarker.

[0218] 53. The test device according to clause 52, wherein the second site comprises two test lines comprising binders immobilised therein for visually determining the presence of at least three biomarkers at two or more different concentrations for each biomarker, wherein the first test line provides a visual determination of the presence of each biomarker at a first concentration for each biomarker and the second test line provides a visual determination of the presence of each biomarker at a second concentration for each biomarker.

[0219] 54. The test device according to clause 53, wherein the second site comprises two test lines comprising binders immobilised therein for visually determining the presence of at least three biomarkers at two or more different concentrations for each biomarker, wherein the first test line provides a visual determination of the presence of each biomarker at a low concentration for each biomarker and the second test line provides a visual determination of the presence of each biomarker at a high concentration for each biomarker.

[0220] 55. The test device according to clause 52-54, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal, bacterial and viral biomarker in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample and wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL or below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above or about 0.50 ng / mL or above in a blood sample. 56. The test device according to clause 52-54, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal, bacterial and viral biomarker in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample and wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above in a blood sample.

[0221] 57. The test device according to clause 52-54, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal, bacterial and viral biomarker in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample and wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.50 ng / mL or above in a blood sample.

[0222] 58. The test device according to clause 52, wherein the second site comprises three test lines comprising binders immobilised therein for visually determining the presence of at least three biomarkers at three or more different concentrations for each biomarker, wherein the first test line provides a visual determination of the presence of each biomarker at a first concentration for each biomarker, the second test line provides a visual determination of the presence of each biomarker at a second concentration for each biomarker, and the third test line provides a visual determination of the presence of each biomarker at a third concentration for each biomarker. 59. The test device according to clause 52, wherein the second site comprises three test lines comprising binders immobilised therein for visually determining the presence of at least three biomarkers at three or more different concentrations for each biomarker, wherein the first test line provides a visual determination of the presence of each biomarker at a low concentration for each biomarker, the second test line provides a visual determination of the presence of each biomarker at a medium concentration for each biomarker, and the third test line provides a visual determination of the presence of each biomarker at a high concentration for each biomarker.

[0223] 60. The test device according to clause 58 or 59, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the fungal, viral and bacterial biomarker substances in the sample each at three different concentrations, wherein the fungal biomarker comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample, the second test line provides a visual determination of BDG corresponding to a concentration of about 81-500 pg / mL in a blood sample and the third test line provides a visual determination of BDG corresponding to a concentration of above about 500 pg / mL in a blood sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample, the second test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60-100 ng / mL in a blood sample and the third test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 100 ng / mL in a blood sample, and wherein the bacterial biomarker substance comprises PCT, wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample, the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25-0.49 ng / mL in a blood sample and the third test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.5 ng / mL or above in a blood sample.

[0224] 61 . The test device according to clause 22, wherein the second site comprises two or more test lines comprising binders immobilised therein for visually determining the presence of at least four biomarkers at two or more different concentrations for each biomarker, wherein each test line provides a visual determination of the presence of each biomarker at two or more different concentrations of each biomarker.

[0225] 62. The test device according to clause 61 , wherein the second site comprises two test lines comprising binders immobilised therein for visually determining the presence of at least four biomarkers at two or more different concentrations for each biomarker, wherein the first test line provides a visual determination of the presence of each biomarker at a first concentration for each biomarker and the second test line provides a visual determination of the presence of each biomarker at a second concentration for each biomarker. 63. The test device according to clause 62, wherein the second site comprises two test lines comprising binders immobilised therein for visually determining the presence of at least four biomarkers at two or more different concentrations for each biomarker, wherein the first test line provides a visual determination of the presence of each biomarker at a low concentration for each biomarker and the second test line provides a visual determination of the presence of each biomarker at a high concentration for each biomarker.

[0226] 64. The test device according to any one of clauses 61-63, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance, a third binder for a bacterial biomarker substance and a fourth binder for a non-specific inflammatory biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal, bacterial, viral and non-specific inflammatory biomarker substance in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample, wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL or below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above or 0.50 ng / mL or above in a blood sample, and wherein the non-specific inflammatory marker comprises C-reactive protein (CRP), and wherein the first test line provides a visual determination of the presence of CRP corresponding to a concentration of below about 9.9 mg / dL in a blood sample and the second test line provides a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

[0227] 65. The test device according to any one of clauses 61-63, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance, a third binder for a bacterial biomarker substance and a fourth binder for a non-specific inflammatory biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal, bacterial, viral and non-specific inflammatory biomarker substance in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample, wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above in a blood sample, and wherein the non-specific inflammatory marker comprises C-reactive protein (CRP), and wherein the first test line provides a visual determination of the presence of CRP corresponding to a concentration of below about 9.9 mg / dL in a blood sample and the second test line provides a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

[0228] 66. The test device according to any one of clauses 61-63, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance, a third binder for a bacterial biomarker substance and a fourth binder for a non-specific inflammatory biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal, bacterial, viral and non-specific inflammatory biomarker substance in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample, wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.50 ng / mL or above in a blood sample, and wherein the non-specific inflammatory marker is C- reactive protein (CRP), and wherein the first test line provides a visual determination of the presence of CRP corresponding to a concentration of below about 9.9 mg / dL in a blood sample and the second test line provides a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

[0229] 67. The test device according to clause 22, wherein the second site comprises three or more test lines comprising binders immobilised therein for visually determining the presence of at least four biomarkers at three or more different concentrations for each biomarker, wherein each test line provides a visual determination of the presence of each biomarker at three or more different concentrations of each biomarker.

[0230] 68. The test device according to clause 67, wherein the second site comprises three test lines comprising binders immobilised therein for visually determining the presence of at least four biomarkers at three or more different concentrations for each biomarker, wherein the first test line provides a visual determination of the presence of each biomarker at a first concentration for each biomarker, the second test line provides a visual determination of the presence of each biomarker at a second concentration for each biomarker and the third test line provides a visual determination of the presence of each biomarker at a third concentration for each biomarker.

[0231] 69. The test device according to clause 68, wherein the second site comprises three test lines comprising binders immobilised therein for visually determining the presence of at least four biomarkers at three or more different concentrations for each biomarker, wherein the first test line provides a visual determination of the presence of each biomarker at a low concentration for each biomarker, the second test line provides a visual determination of the presence of each biomarker at a medium concentration for each biomarker and the third test line provides a visual determination of the presence of each biomarker at a high concentration for each biomarker.

[0232] 70. The test device according to clauses 67-69, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance, a third binder for a bacterial biomarker substance and a fourth binder for a non-specific inflammatory biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the fungal, viral, bacterial and non-inflammatory biomarker substances in the sample each at three different concentrations, wherein the fungal biomarker comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample, the second test line provides a visual determination of BDG corresponding to a concentration of about 81-500 pg / mL in a blood sample and the third test line provides a visual determination of BDG corresponding to a concentration of above about 500 pg / mL in a blood sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample, the second test line provides a visual determination of the presence of MxA corresponding to a concentration of about 61-100 ng / mL in a blood sample and the third test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 100 ng / mL in a blood sample, wherein the bacterial biomarker substance comprises PCT, wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample, the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25-0.49 ng / mL in a blood sample and the third test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.5 ng / mL or above in a blood sample, and wherein the non-specific inflammatory biomarker substance comprises CRP, wherein the first test line provides a visual determination of the presence of CRP corresponding to a concentration of below about 2.0 mg / dL in a blood sample, the second test line provides a visual determination of the presence of CRP corresponding to a concentration of about 2.1-9.9 mg / dL in a blood sample and the third test line provides a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

[0233] 71 . The test device according to any one of the preceding clauses wherein the liquid sample comprises a blood sample. 72. The test device according to any preceding clause wherein the permeable material is planar and the second site is laterally spaced from the first site along the planar permeable material such that the test device provides a lateral flow test device.

[0234] 73. The test device according to any preceding clause, wherein the test device detects two or more biomarkers outlined in Table 1 .

[0235]

[0236]

[0237]

[0238]

[0239]

[0240] Summary of the Figures

[0241] Embodiments and experiments illustrating the principles of the invention will now be discussed with reference to the accompanying figures in which:

[0242] Figure 1a shows a perspective view of an example lateral flow device embodiment which detects four biomarkers on a single test channel, each at three different concentrations (low, medium and high concentrations).

[0243] Figure 1b shows a perspective view of the internal test features of the example device of figure 1a.

[0244] Figures 1c-1g show a perspective view of test results for the example device of figure 1 a.

[0245] Figure 2a shows a perspective view of an example lateral flow device embodiment which detects three biomarkers on a single test channel, each at three different concentrations (low, medium and high concentrations).

[0246] Figures 2b-2e show a perspective view of test results for the example device of figure 2a.

[0247] Figure 3a shows a perspective view of an example lateral flow device embodiment which detects two biomarkers on a single test channel, each at three different concentrations (low, medium and high concentrations).

[0248] Figures 3b-3g show a perspective view of test results for the example device of figure 3a.

[0249] Figure 4a shows a perspective view of an example lateral flow device embodiment which detects four biomarkers on a single test channel, each at two different concentrations (low and high concentrations).

[0250] Figures 4b-4e show a perspective view of test results for the example device of figure 4a. Figure 5a shows a perspective view of an example lateral flow device embodiment which detects three biomarkers on a single test channel, each at two different concentrations (low and high concentrations).

[0251] Figures 5b-5e show a perspective view of test results for the example device of figure 5a.

[0252] Figure 6a shows a perspective view of an example lateral flow device embodiment which detects two biomarkers on a single test channel, each at two different concentrations (low and high concentrations).

[0253] Figure 6b shows a perspective view of test results for the example device of figure 6a.

[0254] Figure 7a shows a perspective view of an example lateral flow device embodiment which detects two biomarkers on a single test channel, wherein one biomarker is tested at a single concentration and the second biomarker is detected at two different concentrations (low and high concentrations).

[0255] Figures 7b-7f show a perspective view of test results for the example device of figure 7a.

[0256] Figures 8a and 8b show a perspective view of test results for an example lateral flow device embodiment which detects two biomarkers, where each biomarker is tested on a different test channel, wherein both biomarkers are tested at three different concentrations (low, medium and high concentrations) respectively.

[0257] Figures 9a-9c show a perspective view of test results for an example lateral flow device embodiment which detects three biomarkers, where each biomarker is tested on a different test channel, wherein all three biomarkers are tested at three different concentrations (low, medium and high concentrations) respectively.

[0258] Figures 10a-10c show a perspective view of test results for an example lateral flow device embodiment which detects four biomarkers, where each biomarker is tested on a different test channel, wherein all four biomarkers are tested at three different concentrations (low, medium and high concentrations) respectively.

[0259] Figures 11 a and 11 b show a perspective view of test results for an example lateral flow device embodiment which detects two biomarkers, where each biomarker is tested on a different test channel, wherein both biomarkers are tested at two different concentrations (low and high concentrations) respectively.

[0260] Figures 12a-12c show a perspective view of test results for an example lateral flow device embodiment which detects three biomarkers, where each biomarker is tested on a different test channel, wherein all three biomarkers are tested at two different concentrations (low and high concentrations) respectively.

[0261] Figures 13a-13c show a perspective view of test results for an example lateral flow device embodiment which detects four biomarkers, where each biomarker is tested on a different test channel, wherein all four biomarkers are tested at two different concentrations (low and high concentrations) respectively.

[0262] Figures 14a-14c show a perspective view of test results for an example lateral flow device embodiment which detects two biomarkers, where each biomarker is tested on a different test channel, wherein one biomarker is tested at a single threshold concentration and the other biomarker is tested at two different concentrations (low and high concentrations).

[0263] Figures 15a-15c show a perspective view of test results for an example lateral flow device embodiment which detects two biomarkers, where each biomarker is tested on a different test channel, wherein one biomarker is tested at a single threshold concentration and the other biomarker is tested at three different concentrations (low, medium and high concentrations).

[0264] Detailed Description of the Invention

[0265] Aspects and embodiments of the present invention will now be discussed with reference to the accompanying figures. Further aspects and embodiments will be apparent to those skilled in the art. All documents mentioned in this text are incorporated herein by reference.

[0266] Referring to figures 1a, 2a, 3a, 4a, 5a, 6a and 7a, there is shown an example embodiment of the current invention in the form of a housed lateral flow device (101), which is unhoused in figure 1 b to show the internal test features and construction.

[0267] A test device (101) is configured as a lateral flow device for detecting the presence of biomarkers in a liquid sample. The device comprises a permeable material in the form of porous pads (105, 106, 107) defining: a first portion (105, 106) comprising a sample pad (105) and a conjugate pad (106) collectively configured for providing a first site for application of the liquid sample and for conjugates movably supported therein wherein each conjugate comprises a binder for a respective biomarker coupled to a respective detector reagent; and, a second portion (107) defining a test zone and configured relative to the first portion (105, 106) so as to permit capillary flow communication therebetween. The second portion (107) provides a plurality of separate second sites (114-125) each of which is spaced from the first site (105, 106) for visually determining the presence of the detector reagent (coloured particle). Each one of the second sites (114- 125) comprises a binder immobilized therein which binds to a respective biomarker either at a single concentration, at two concentrations or at three concentrations of the biomarker.

[0268] The first portion comprises two separate component parts (two porous pads, 105, 106) that divide the first site into two contiguous sub-sites (105, 106) in mutual contact with each other to support a capillary flow of liquid sample between them. Application of the liquid sample occurs via a sample port (102) exposing a location upon the porous sample pad (105) forming a first sub-site that is laterally spaced from, and in flow communication with, the location of the conjugates in the porous conjugate pad (106) forming a second sub-site. The second component part (106) defines a so-called “conjugate pad” in that it holds the awaiting biomarker binders (e.g., conjugates).

[0269] Thus, in this example, the first component part (105) of the first portion defines a first of the two sub-sites and comprises a porous sample pad for receiving the liquid sample and for supporting capillary fluid flow of the received sample to the second component part (106) of the first portion. The second component part forms a second one of the two sub-sites that is in capillary fluid flow communication with the first component part and contains the conjugates movably supported therein. When the liquid sample comprises a blood sample, the first component part (105) serves to remove / filter red blood cells from the sample during capillary flow of the sample thorough it. This means that the remaining parts of the sample delivered to the second component part (106) are substantially free of red blood cells.

[0270] In other examples (not shown), the first portion (105, 106) may comprise one continuous porous pad defining the whole of the first site for application of the liquid sample and for conjugates movably supported therein.

[0271] The first portion (105, 106) comprises, for a first biomarker, a first moveably supported conjugate comprising a first binder for a fungal, viral or bacterial biomarker substance coupled to a first coloured particle, and for at least one additional biomarker selected from a fungal, viral or bacterial biomarker substance, an additional moveably supported conjugate comprising a binder for such fungal, viral or bacterial biomarker substance coupled to an additional coloured particle The first, additional and further detector reagents (coloured particles) may have different respective colours easily distinguishable from each other. The detector reagents (coloured particles) may have respective colours easily distinguishable from the colour of the second portion (107) where they are to be immobilized, bound and accumulated in use.

[0272] At the rear of the device, a wick (108) is located, which is typically a porous pad and serves only to mop up excess reagents and prevent the device from leaking once the test has run.

[0273] Example 1

[0274] The Lateral flow device (101) consists of an outer housing (104) which may be made of a suitable polymeric material in a simple snap together configuration. At one end of the device and running through the top surface of the outer housing (104) is a sample port (102), which provides an access point forthe sample to enter the device. The size and shape of the sample port (102) may be such that it helps to meter the volume of the sample that can be introduced. Directly below the sample port (102) is the sample pad (105) which serves to receive the sample and filter / hold back any unwanted sample components that may negatively affect the test (or how the test results are observed).

[0275] The material requirements for such a sample pad will depend on the sample type, but an example for whole blood samples may be (but not limited to) “Whatman 5” 2.5pm filter paper, which can effectively retain red blood cells and prevent them from inhibiting the visual appraisal of the test results. Adjacent to and in direct contact and fluid communication with the sample pad (105) is the conjugate pad (106) which holds the test antibody conjugate reagents, including the control line conjugates. The concentrations of which are pre-determined based on the required clinical threshold values.

[0276] Adjacent to and in direct contact and fluid communication with the conjugate pad (106) is the test zone (107) made from a membrane of a suitable material known to those skilled in the art, for example (but not limited to) nitrocellulose (e.g., Sartorius CN95). The test zone (107) has distinctive test lines (114-125) which have specific individual capture antibodies immobilised on each line. The individual capture antibodies are specific for a particular biomarker as described herein.

[0277] In Figures 1 a, and 1c-1g, four biomarkers are being tested on a single channel, wherein each biomarker is being tested at three different concentrations (low, medium and high concentrations).

[0278] In Figures 1 c-3g, test lines 1-3 (114-116) are specific to capturing the BDG-conjugate complex only, where test line 1 (114) has specific immobilised capture antibodies ready to capture BDG-conjugate complexes corresponding to a concentration of 60-80 pg / mL (i.e. a low concentration) in an (e.g. undiluted) blood sample, test line 2 (115) has specific immobilised capture antibodies capture BDG- conjugate complexes corresponding to a concentration of 81-500 pg / mL (i.e. a medium concentration) in an (e.g. undiluted) blood sample and test line 3 (116) has specific immobilised capture antibodies ready to capture BDG-conjugate complexes corresponding to a concentration of above 501 pg / mL (i.e. a high concentration) in an (undiluted) blood sample.

[0279] In Figures 1 c-3g, test lines 4-6 (117-119) are specific to capturing the PCT-conjugate complex only, wherein test line 4 (117) has specific immobilised capture antibodies ready to capture PCT-conjugate complexes corresponding to a concentration of 0.1-0.24 ng / mL (i.e. a low concentration) in an (e.g. undiluted) blood sample, test line 5 (118) has specific immobilised capture antibodies capture PCT- conjugate complexes corresponding to a concentration of 0.25-0.49 ng / mL (i.e. a medium concentration) in an (e.g. undiluted) blood sample and test line 6 (1 19) has capture PCT-conjugate complexes corresponding to a concentration of above 0.5 ng / mL (i.e. a high concentration) in an (e.g. undiluted) blood sample.

[0280] In Figures 1 c-3g, test lines 7-9 (120-122) in this example will be specific to capturing the MxA-conjugate complex only, wherein test line 7 (120) has specific immobilised capture antibodies ready to capture MxA- conjugate complexes corresponding to a concentration of 40-60 ng / mL (i.e. a low concentration) in an (e.g. undiluted) blood sample, test line 8 (121) has specific immobilised capture antibodies capture MxA- conjugate complexes corresponding to a concentration of 80-100 ng / mL (i.e. a medium concentration) in an (e.g. undiluted) blood sample and test line 9 (122) has specific immobilised capture antibodies ready to capture MxA-conjugate complexes corresponding to a concentration of above 101 ng / mL (i.e. a high concentration) in an (e.g. undiluted) blood sample.

[0281] In Figures 1 c-3g, test lines 10-12 (123-125) are specific to capturing the CRP-conjugate complex only, wherein test line 10 (123) has specific immobilised capture antibodies ready to capture CRP-conjugate complexes corresponding to a concentration of 1 .0-2.0 mg / dL (i.e. a low concentration) in an (undiluted) blood sample, test line 11 (124) has specific immobilised capture antibodies capture CRP-conjugate complexes corresponding to a concentration of 2.1-9.9 mg / dL (i.e. a medium concentration) in an (undiluted) blood sample and test line 12 (125) has specific immobilised capture antibodies ready to capture CRP-conjugate complexes corresponding to a concentration of above 10.0 mg / dL (i.e. a high concentration) in an (undiluted) blood sample.

[0282] In Figure 1 c, the test lines are configured in a configuration wherein the high concentration test line (116, 119, 122, 125) for each biomarker appears closest to the control line and the lowest concentration test line (114, 117, 120 and 123) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, test lines 1-3 (114-116) are positive, as shown by a solid test line, suggesting a high (above 501 pg / ml) concentration of BDG in an (e.g. undiluted) blood sample, test lines 4-6 (117- 119) are positive, as shown by a solid test line, suggesting a high (above 0.5 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample, test lines 7-9 (120-122) are positive, as shown by a solid test line, suggesting a high (above 101 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample and test lines 10-12 (123-125) are positive, as shown by a solid test line, suggesting a high (above 10.0 mg / dL) concentration of CRP in an (e.g. undiluted) blood sample. This suggests that BDG, PCT, MxA and CRP are all present in the sample at a high concentration and therefore a bacterial, fungal and viral infection is detectable along with high levels of inflammation. The clinician may therefore advise that the patient has a severe fungal, bacterial and viral co-infection present, whereby the fungal, bacterial and viral infections are present to significant levels that would warrant the urgent prescribing of an antifungal, antibiotic and antiviral agent. The high level of each infection type and the high level of inflammation suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0283] In Figure 1d, the test lines are configured in an alternative configuration wherein the high concentration test line (116, 119, 122, 125) for each biomarker appears closest to the sample port and the lowest concentration test line (114, 117, 120 and 123) for each biomarker appears closest to the control line for each sub-set of test lines. In this example, test lines 1 and 2 (114 and 115) are positive, as shown by a solid test line, but test line 3 (116) is negative, as there is an absence of any visual solid test line. This suggests a medium (81-500 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample. Test lines 4 and 5 (117 and 118 ) are positive, as shown by a solid test line, but test line 6 (119) is negative, as there is an absence of any visual solid test line. This suggests a medium (0.25-0.49 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test lines 7 and 8 (120 and 121) are positive, as shown by a solid test line, but test line 9 (122) is negative, as there is an absence of any visual solid line. This suggests a medium (80-100 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. Test lines 10 and 11 (123 and 124) are positive, as shown by a solid test line, but test line 12 (125) is negative, as there is an absence of any visual solid line. This suggests a medium (2.1-9.9 mg / dL) concentration of CRP in an (e.g. undiluted) blood sample. This suggests that BDG, PCT, MxA and CRP are all present in the sample at a medium concentration and therefore a bacterial, fungal and viral infection is detectable along with medium levels of inflammation. The clinician may therefore advise that the patient has a fungal, bacterial and viral co-infection present, whereby the fungal, bacterial and viral infections are present to significant levels that would warrant the prescribing of an antifungal, antibiotic and antiviral agent. The medium level of each infection and level of inflammation suggests that if not already in hospital, the patient should be admitted for treatment and monitored as a potential sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitorthe effectiveness of any administered treatment. In Figure 1 e, the test lines are configured in an alternative configuration wherein the high concentration test line (116, 119, 122, 125) for each biomarker appears closest to the sample port and the lowest concentration test line (114, 117, 120 and 123) for each biomarker appears closest to the control line for each sub-set of test lines. In this example, test line 1 (114) is positive, as shown by a solid test line, but test lines 2 and 3 (115 and 116) are negative, as there is an absence of any visual solid test line. This suggests a low (60-80 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample. Test line 4 (117) is positive, as shown by a solid test line, but test lines 5 and 6 (118 and 119) are negative, as there is an absence of any visual solid test line. This suggests a low (0.1-0.24 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test line 7 (120) is positive, as shown by a solid test line, but test lines 8 and 9 (121 and 122) are negative, as there is an absence of any visual solid line. This suggests a low (40-60 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. Test line 10 (123) is positive, as shown by a solid test line, but test lines 11 and 12 (124 and 125) are negative, as there is an absence of any visual solid line. This suggests a low (1.0-2.0 mg / dL) concentration of CRP in an (e.g. undiluted) blood sample. This suggests that BDG, PCT, MxA and CRP are all present in the sample at a low concentration and therefore a bacterial, fungal and viral infection is detectable along with low levels of inflammation. The clinician may therefore advise that the patient has a low level fungal, bacterial and viral co-infection. The patient should be monitored and clinical judgement used to determine if antimicrobial agents are required at this stage as the levels of infection and inflammation are still low and there is a very low risk of sepsis. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0284] In Figure 1f, the test lines are configured in a configuration wherein the high concentration test line (116, 119, 122, 125) for each biomarker appears closest to the control line and the lowest concentration test line (114, 117, 120 and 123) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, all test lines are negative for all biomarkers, as there is an absence of any visual solid line. This suggests that BDG, PCT, MxA and CRP are not detectable in the sample, and therefore a bacterial, fungal or viral infection and inflammation can be ruled out. The clinician may therefore advise that as the patient’s sample shows no abnormal levels of these biomarkers, that there is no evidence of infection or inflammation and therefore antimicrobial treatment is not required, and sepsis is not suspected.

[0285] In Figure 1g, the test lines are configured in a configuration wherein the high concentration test line (116, 119, 122, 125) for each biomarker appears closest to the sample port and the lowest concentration test line (114, 117, 120 and 123) for each biomarker appears closest to the control line for each sub-set of test lines. In this example, test lines 1-3 (114-116) for BDG are positive, as shown by a visual solid test line, suggesting a high (above 501 pg / ml) concentration of BDG in an (e.g. undiluted) blood sample. Test line 4 (117) is positive, as shown by a solid test line, but test lines 5 and 6 (118 and 119) are negative, as there is an absence of any visual solid test line. This suggests a low (of 0.1-0.24 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test lines 7 and 8 (120 and 121) are positive, as shown by a solid test line, but test line 9 (122) is negative, as there is an absence of any visual solid line. This suggests a medium (80-100 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. Test lines 10-12 are all negative as there is an absence of any visual solid line. This suggests that CRP is not detectable in the sample. This suggests that BDG is present in a high concentration in the sample, PCT is present in a low concentration in the sample and MxA is present in a medium concentration in the sample, and that no inflammation is present. The clinician may therefore advise that the patient has a fungal, bacterial and viral co-infection, where the fungal and viral infections are present to a significant level that may warrant the prescribing of an antifungal and an antiviral agent. Given the lack of inflammation present there is no urgent requirement for treatment and sepsis is not suspected at this moment in time. The clinician may choose to monitor and retest the patient again after a defined period of time to determine if the patient’s condition has changed to an extent warranting any further action. As the bacterial infection is present to a low level the patient may be monitored and clinical judgement used to determine if appropriate antibiotics are required at this stage as the level of infection is still low and there is a very low risk of sepsis. In Figures 1 c-1 g, there is a solid visual control line (112) indicating that the test has run correctly and so is valid (e.g., the results are valid).

[0286] In Figures 2a-2e, three biomarkers are being tested on a single channel, wherein each biomarker is being tested at three different concentrations (low, medium and high concentrations).

[0287] In Figure 2b, the test lines are configured in a configuration wherein the high concentration test line (116, 119, 122,) for each biomarker appears closest to the control line and the lowest concentration test line (114, 117, 120) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, test lines 1-3 (114-116) are positive, as shown by a solid test line, suggesting a high (above 501 pg / ml) concentration of BDG in an (e.g. undiluted) blood sample, test lines 4-6 (117-119) are positive, as shown by a solid test line, suggesting a high (above 0.5 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample, test lines 7-9 (120-122) are positive, as shown by a solid test line, suggesting a high (above 101 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. This suggests that BDG, PCT and MxA are all present in the sample at a high concentration and therefore a bacterial, fungal and viral infection is detectable. The clinician may therefore advise that the patient has a severe fungal, bacterial and viral co-infection present, whereby the fungal, bacterial and viral infections are present to significant levels that would warrant the urgent prescribing of an antifungal, antibiotic and antiviral agent. The high level of each infection type suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0288] In Figure 2c, the test lines are configured in an alternative configuration wherein the high concentration test line (116, 119, 122) for each biomarker appears closest to the sample port and the lowest concentration test line (114, 117, 120) for each biomarker appears closest to the control line for each subset of test lines. In this example, test lines 1 and 2 (114 and 115) are positive, as shown by a solid test line, but test line 3 (116) is negative, as there is an absence of any visual solid test line. This suggests a medium (81-500 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample. Test lines 4 and 5 (117 and 118 ) are positive, as shown by a solid test line, but test line 6 (119) is negative, as there is an absence of any visual solid test line. This suggests a medium (0.25-0.49 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test lines 7 and 8 (120 and 121) are positive, as shown by a solid test line, but test line 9 (122) is negative, as there is an absence of any visual solid line. This suggests a medium (80-100 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. This suggests that BDG, PCT and MxA are all present in the sample at a medium concentration and therefore a bacterial, fungal and viral infection is detectable. The clinician may therefore advise that the patient has a fungal, bacterial and viral co-infection present, whereby the fungal, bacterial and viral infections are present to significant levels that would warrant the prescribing of an antifungal, antibiotic and antiviral agent. The medium level of each infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as a potential sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0289] In Figure 2d, the test lines are configured in an alternative configuration wherein the high concentration test line (116, 119 and 122) for each biomarker appears closest to the sample port and the lowest concentration test line (114, 117 and 120) for each biomarker appears closest to the control line. In this example, test line 1 (114) is positive, as shown by a solid test line, but test lines 2 and 3 (115 and 116) are negative, as there is an absence of any visual solid test line. This suggests a low (60-80 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample. Test line 4 (1 17) is positive, as shown by a solid test line, but test lines 5 and 6 (118 and 119) are negative, as there is an absence of any visual solid test line. This suggests a low (of 0.1-0.24 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test line 7 (120) is positive, as shown by a solid test line, but test lines 8 and 9 (121 and 122) are negative, as there is an absence of any visual solid line. This suggests a low (40-60 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. This suggests that BDG, PCT and MxA are all present in the sample at a low concentration and therefore a bacterial, fungal and viral infection is detectable at a low concentration. The clinician may therefore advise that the patient has a low level fungal, bacterial and viral co-infection. The patient should be monitored and clinical judgement used to determine if antimicrobial agents are required at this stage as the levels of infection are still low and there is a very low risk of sepsis. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0290] In Figure 2e, the test lines are configured in a configuration wherein the high concentration test line (116, 119 and 122) for each biomarker appears closest to the sample port and the lowest concentration test line (114, 117 and 120) for each biomarker appears closest to the control line for each sub-set of test lines. In this example, test lines 1 (114) for BDG is positive, as shown by a visual solid test line, but test lines 2 and 3 (115 and 116) are negative, as there is an absence of any visual solid line. This suggests a low (60-80 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample. Test lines 4 and 5 (117 and 118) are positive, as shown by a solid test line, but test line 6 (119) is negative, as there is an absence of any visual solid test line. This suggests a medium (0.25-0.49 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test lines 7, 8 and 9 (120, 121 and 122) are positive, as shown by a solid test line. This suggests a high (above 101 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. This suggests that BDG is present in a low concentration in the sample, PCT is present in a medium concentration in the sample and MxA is present in a high concentration in the sample. The clinician may therefore advise that the patient has a fungal, bacterial and viral co-infection present, whereby the viral and bacterial infections are present to levels that would warrant the prescribing of an antiviral and an antibacterial agent. The significant level of each infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The patient should be monitored and clinical judgement used to determine if antifungal agents are required at this stage as the level of fungal infection is still low. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitorthe effectiveness of any administered treatment.

[0291] In Figures 2b-2e, there is a solid visual control line (112) indicating that the test has run correctly and so is valid (e.g., the results are valid).

[0292] In Figures 3a-3e, two biomarkers are being tested on a single channel. In Figures 3a and 3b, each biomarker is being tested at three different concentrations (low, medium and high concentrations). In Figures 3c-3e, one biomarker is tested at a single threshold concentration and the other biomarker is tested at three different concentrations (low, medium and high concentration). In Figures 3c-3e, test line 1 (114) detects BDG in the first test channel and is specific to capturing the BDG-conjugate complex only, where test line 1 (114) has specific immobilised capture antibodies ready to capture BDG-conjugate complexes at a single threshold level corresponding to a concentration of 80 pg / mL in an (e.g. undiluted) blood sample.

[0293] In Figure 3b, the test lines are configured in a configuration wherein the high concentration test line (116, 119) for each biomarker appears closest to the control line and the lowest concentration test line (114, 117) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, test lines 1-3 (114-116) are positive, as shown by a solid test line, suggesting a high (above 501 pg / ml) concentration of BDG in an (e.g. undiluted) blood sample and test lines 4-6 (117-119) are positive, as shown by a solid test line, suggesting a high (above 0.5 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. This suggests that BDG and PCT are both present in the sample at a high concentration and therefore a bacterial and fungal infection is detectable. The clinician may therefore advise that the patient has a fungal and bacterial co-infection present, whereby the fungal and bacterial infections are present to significant levels that would warrant the prescribing of an antifungal and antibiotic agent. The high level of each infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitorthe effectiveness of any administered treatment. In Figure 3c, BDG is tested at a single threshold concentration and PCT is tested at three concentrations (low, medium and high). The test lines for PCT are configured in a configuration wherein the high concentration test line (119) appears closest to the control line and the lowest concentration test line (117) appears closest to the sample port. In this example, only one test line is present for BDG which is positive as shown by a solid test line, suggesting the presence of BDG at the single threshold concentration (e.g. 80 pg / mL) in an (e.g. undiluted) blood sample. Test lines 2 and 3 for BDG are not included within this arrangement device. Test lines 4 and 5 (117 and 118) for PCT are positive, as shown by a solid test line, but test line 6 (119) is negative as there is an absence of any visual solid test line. This suggests a medium (0.25-0.49 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. This suggests that BDG is present at a desired threshold level and PCT is present at a medium concentration. Therefore, a bacterial and fungal infection are detectable in the sample. The clinician may therefore advise that the patient has a fungal and bacterial co-infection present, whereby the fungal and bacterial infections are present to levels that may warrant the prescribing of an antifungal and an antibiotic agent. The medium level of the bacterial infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as a potential sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0294] In Figure 3d, BDG is tested at a single threshold concentration and PCT is tested at three concentrations (low, medium and high). The test lines for PCT are configured in a configuration wherein the high concentration test line (119) for appears closest to the control line and the lowest concentration test line (117) appears closest to the sample port. In this example, only one test line is present for BDG which is positive as shown by a solid test line, suggesting the presence of BDG at the single threshold concentration (e.g. 80 pg / mL) in an (e.g. undiluted) blood sample. Test lines 2 and 3 for BDG are not included within this arrangement device. Test lines 4 (1 17) for PCT is positive, as shown by a solid test line, but test lines 5 and 6 (118 and 119) are negative as there is an absence of any visual solid test line. This suggests a low (0.1-0.24 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. This suggests that BDG is present at a desired threshold level and PCT is present at a low concentration. Therefore, a bacterial and fungal infection may be detectable in the sample. The clinician may therefore advise that the patient has a fungal and bacterial co-infection wherein the fungal infection is present to a level that an antifungal agent may be prescribed. The patient should be monitored, and clinical judgement used as the infection may be significant enough that it may progress towards sepsis. As the bacterial infection is present to a low level the patient should be monitored and clinical judgement used to determine if an antibiotic agent is required at this stage as the level of infection is still low and there is only a low risk of sepsis. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitorthe effectiveness of any administered treatment.

[0295] In Figure 3e, BDG is tested at a single threshold concentration and PCT is tested at three concentrations (low, medium and high). The test lines for PCT are configured in a configuration wherein the high concentration test line (119) for appears closest to the control line and the lowest concentration test line (117) appears closest to the sample port. In this example, only one test line is present for BDG which is positive as shown by a solid test line, suggesting the presence of BDG at the single threshold concentration (e.g. 80 pg / mL) in an (e.g. undiluted) blood sample. Test lines 2 and 3 for BDG are not included within this arrangement device. Test lines 4-6 (117-119) for PCT are negative as there is an absence of any visual solid test line. This suggests the absence of PCT in an (e.g. undiluted) blood sample. This suggests that BDG is present at a desired threshold level and PCT is not present within the sample. Therefore, a fungal infection may be detectable in the sample. The clinician may therefore advise that the patient has a fungal infection wherein the fungal infection is present to a level that an antifungal agent may be prescribed. The patient should be monitored, and clinical judgement used as the infection may be significant enough that it may progress towards sepsis. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0296] Figure 3f shows the same results as Figure 3c and Figure 3g shows the same results as Figure 3d, but the three test lines of PCT (low / medium / high) are configured the opposite way round compared to the test lines of Figures 3f and 3g. In Figure 3f and Figure 3g, the high concentration test line (1 19) for PCT appears closest to the sample port and the lowest concentration test line (117) appears closest to the control line.

[0297] In Figures 3b-3g, there is a solid visual control line (112) indicating that the test has run correctly and so is valid (e.g., the results are valid).

[0298] Example 2

[0299] In Example 2, the lateral flow device has the same configuration as the lateral flow device used in Example 1 .

[0300] In Figures 4b-7f, test lines 1 and 2 (214 and 215) are specific to capturing the BDG-conjugate complex only, where test line 1 (214) has specific immobilised capture antibodies ready to capture BDG-conjugate complexes corresponding to a concentration of 60-80 pg / mL (i.e. a low concentration) in an (e.g. undiluted) blood sample and test line 2 (215) has specific immobilised capture antibodies capture BDG- conjugate complexes corresponding to a concentration of above 81 pg / mL (i.e. a high concentration) in an (e.g. undiluted) blood sample.

[0301] In Figures 4b-7f, test lines 3 and 4 (217 and 218) are specific to capturing the PCT-conjugate complex only, wherein test line 3 (217) has specific immobilised capture antibodies ready to capture PCT- conjugate complexes corresponding to a concentration of 0.1-0.24 ng / mL (i.e. a low concentration) in an (e.g. undiluted) blood sample and test line 4 (218) has specific immobilised capture antibodies capture PCT-conjugate complexes corresponding to a concentration of 0.25 ng / mL or above (i.e. a high concentration). In Figures 4b-7f, test lines 5 and 6 (220 and 221) are specific to capturing the MxA-conjugate complex only, wherein test line 5 (220) has specific immobilised capture antibodies ready to capture MxA- conjugate complexes corresponding to a concentration of 40-60 ng / mL (i.e. a low concentration) in an (e.g. undiluted) blood sample and test line 6 (221) has specific immobilised capture antibodies capture MxA-conjugate complexes corresponding to a concentration of 80 ng / mL or above (i.e. a high concentration).

[0302] In Figures 4b-7f, test lines 7 and 8 (223 and 224) are specific to capturing the CRP-conjugate complex only, wherein test line 7 (223) has specific immobilised capture antibodies ready to capture CRP- conjugate complexes corresponding to a concentration of 1 .0-9.9 mg / dL (i.e. a low concentration) in an (undiluted) blood sample and test line 8 (224) has specific immobilised capture antibodies capture CRP- conjugate complexes corresponding to a concentration of 10.0 mg / dL or above (i.e. a high concentration).

[0303] In Figures 4a-4e, four biomarkers are being tested on a single channel within the device, wherein each biomarker is being tested at two different concentrations (low and high concentrations).

[0304] In Figure 4b, the test lines are configured in a configuration wherein the high concentration test line (215 218, 221 , 224) for each biomarker appears closest to the control line and the lowest concentration test line (214, 217, 220 and 223) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, test lines 1 and 2 (214 and 215) for BDG are positive, as shown by a solid test line, suggesting a high (above 81 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample, test lines 3 and 4 (217 and 218) are positive, as shown by a solid test line, suggesting a high (0.25 ng / mL or above) concentration of PCT in an (e.g. undiluted) blood sample, test lines 5 and 6 (220 and 221) are positive, as shown by a solid test line, suggesting a high (80 ng / mL or above) concentration of MxA in an (e.g. undiluted) blood sample and test lines 7 and 8 (223 and 224) are positive, as shown by a solid test line, suggesting a high (10.0 mg / dL or above) concentration of CRP in an (e.g. undiluted) blood sample. This suggests that BDG, PCT, MxA and CRP are all present in the sample at a high concentration and therefore a bacterial, fungal and viral infection is detectable along with high levels of inflammation. The clinician may therefore advise that the patient has a severe fungal, bacterial and viral co-infection present, whereby the fungal, bacterial and viral infections are present to significant levels that would warrant the urgent prescribing of an antifungal, antibiotic and antiviral agent. The high levels of each infection type and the high level of inflammation suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0305] In Figure 4c, the test lines are configured in a configuration wherein the high concentration test line (215 218, 221 , 224) for each biomarker appears closest to the sample port and the lowest concentration test line (214, 217, 220 and 223) for each biomarker appears closest to the control line for each sub-set of test lines. In this example, test line 1 (214) for BDG is positive, as shown by a solid test line, but test line 2 (215) is negative, as there is an absence of any visual solid test line. This suggests a low (above 60-80 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample. Test line 3 (217) is positive, as shown by a solid test line, but test line 4 (218) is negative, as there is an absence of any visual solid test line. This suggests a low (of 0.1-0.24 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test line 5 (220) is positive, as shown by a solid test line, but test line 6 is negative, as there is an absence of any visual solid test line. This suggests a low (40-60 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. Test line 7 (223) is positive, as shown by a solid test line, but test line 8 (224) is negative, as there is an absence of any visual solid test line. This suggests a low (1 .0-9.9 mg / dL) concentration of CRP in an (e.g. undiluted) blood sample. This suggests that BDG, PCT, MxA and CRP are all present in the sample at a low concentration and therefore a bacterial, fungal and viral infection may be detectable along with low levels of inflammation. The clinician may therefore advise that the patient has a low level fungal, bacterial and viral co-infection. The patient should be monitored and clinical judgement used to determine if antimicrobial agents are required at this stage as the levels of infection and inflammation are still low and there is a very low risk of sepsis. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitorthe effectiveness of any administered treatment.

[0306] In Figure 4d, the test lines are configured in a configuration wherein the high concentration test line (215 218, 221 , 224) for each biomarker appears closest to the control line and the lowest concentration test line (214, 217, 220 and 223) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, all test lines for BDG, PCT, MxA and CRP are negative, as there is an absence of any visual solid test line. This suggests that BDG, PCT, MxA and CRP are not detectable in the sample and therefore a bacterial, fungal and viral infection may not be present, along with no inflammation. The clinician may therefore advise that as the patient’s sample shows no abnormal levels of these biomarkers, that there is no evidence of infection or inflammation and therefore antimicrobial treatment is not required, and sepsis is not suspected.

[0307] In Figure 4e, the test lines are configured in a configuration wherein the high concentration test line (215 218, 221 , 224) for each biomarker appears closest to the sample port and the lowest concentration test line (214, 217, 220 and 223) for each biomarker appears closest to the control line for each sub-set of test lines. In this example, test lines 1 and 2 (214 and 215) for BDG are positive, as shown by a solid test line. This suggests a high (above 81 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample. Test line 3 (217) is positive, as shown by a solid test line, but test line 4 (218) is negative, as there is an absence of any visual solid test line. This suggests a low (of 0.1-0.24 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test lines 5 and 6 (220 and 221) are negative, as there is an absence of any visual solid test line. This suggests that MxA is not detected in the sample. Test lines 7 and 8 (223 and 224) are positive, as shown by a solid test line, suggesting a high (10.0 mg / dL or above) concentration of CRP in an (e.g. undiluted) blood sample. This suggests that BDG and CRP are present in the sample at a high concentration, PCT is present in the sample at a low concentration and MxA is not detectable within the sample. This suggests a fungal and bacterial co-infection with high levels of inflammation. The clinician may therefore advise that the patient has a fungal and bacterial co-infection present, whereby the fungal infection is present to a significant level that would warrant the prescribing of an antifungal agent. As the bacterial infection is present to a low level the patient should be monitored and clinical judgement used to determine if an antibiotic agent is required at this stage as the level of infection is still low. The high level of inflammation suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitorthe effectiveness of any administered treatment.

[0308] In Figures 4b-4e, there is a solid visual control line (212) indicating that the test has run correctly and so is valid (e.g., the results are valid).

[0309] In Figures 5a-5e, three biomarkers are being tested on a single channel within the device, wherein each biomarker is being tested at two different concentrations (low and high concentrations).

[0310] In Figure 5b, the test lines are configured in a configuration wherein the high concentration test line (215 218, 221) for each biomarker appears closest to the control line and the lowest concentration test line (214, 217, 220) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, test lines 1 and 2 (214 and 215) for BDG are positive, as shown by a solid test line, suggesting a high (above 81 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample, test lines 3 and 4 (217 and 218) are positive, as shown by a solid test line, suggesting a high (0.25 ng / mL or above) concentration of PCT in an (e.g. undiluted) blood sample and test lines 5 and 6 (220 and 221) are positive, as shown by a solid test line, suggesting a high (80 ng / mL or above) concentration of MxA in an (e.g. undiluted) blood sample. This suggests that BDG, PCT and MxA are all present in the sample at a high concentration and therefore a bacterial, fungal and viral infection is detectable. The clinician may therefore advise that the patient has a fungal, bacterial and viral co-infection present, whereby the fungal, bacterial and viral infections are present to significant levels that would warrant the prescribing of an antifungal, antibiotic and antiviral agent. The high level of each infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitorthe effectiveness of any administered treatment.

[0311] In Figure 5c, the test lines are configured in a configuration wherein the high concentration test line (215 218, 221) for each biomarker appears closest to the sample port and the lowest concentration test line (214, 217, 220) for each biomarker appears closest to the control line for each sub-set of test lines. In this example, test line 1 (214) for BDG is positive, as shown by a solid test line, but test line 2 (215) is negative, as there is an absence of any visual solid test line. This suggests a low (above 60-80 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample. Test line 3 (217) is positive, as shown by a solid test line, but test line 4 (218) is negative, as there is an absence of any visual solid test line. This suggests a low (of 0.1-0.24 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test line 5 (220) is positive, as shown by a solid test line, but test line 6 is negative, as there is an absence of any visual solid test line. This suggests a low (40-60 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. This suggests that BDG, PCT and MxA are all present in the sample at a low concentration and therefore a bacterial, fungal and viral infection may be detectable. The clinician may therefore advise that the patient has a low level fungal, bacterial and viral co-infection. The patient should be monitored and clinical judgement used to determine if antimicrobial agents are required at this stage as the levels of infection are still low and there is only a low risk of sepsis. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0312] In Figure 5d, the test lines are configured in a configuration wherein the high concentration test line (215 218, 221) for each biomarker appears closest to the control line and the lowest concentration test line (214, 217, 220) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, all test lines for BDG, PCT and MxA are negative, as there is an absence of any visual solid test line. This suggests that BDG, PCT and MxA are not detectable in the sample and therefore a bacterial, fungal and viral infection may not be present. The clinician may therefore advise that as the patient’s sample shows no abnormal levels of these biomarkers, that there is no evidence of infection and therefore antimicrobial treatment is not required, and sepsis is not suspected.

[0313] In Figure 5e, the test lines are configured in a configuration wherein the high concentration test line (215 218, 221) for each biomarker appears closest to the sample port and the lowest concentration test line (214, 217, 220) for each biomarker appears closest to the control line for each sub-set of test lines. In this example, test lines 1 and 2 (214 and 215) for BDG are positive, as shown by a solid test line. This suggests a high (above 81 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample. Test line 3 (217) is positive, as shown by a solid test line, but test line 4 (218) is negative, as there is an absence of any visual solid test line. This suggests a low (of 0.1-0.24 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test lines 5 and 6 (220 and 221) are negative, as there is an absence of any visual solid test line. This suggests that MxA is not detected in the sample. This suggests that BDG is present in the sample at a high concentration, PCT is present in the sample at a low concentration and MxA is not detectable within the sample. This suggests a fungal and bacterial co-infection. The clinician may therefore advise that the patient has a fungal and bacterial co-infection present, whereby the fungal infection is present to a significant level that would warrant the prescribing of an antifungal. The high level of fungal infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. As the bacterial infection is present to a low level the patient should be monitored and clinical judgement used to determine if an antibiotic agent is also required at this stage. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitorthe effectiveness of any administered treatment.

[0314] In Figures 5b-5e, there is a solid visual control line (212) indicating that the test has run correctly and so is valid (e.g., the results are valid).

[0315] In Figures 6a and 6b, two biomarkers are being tested on a single channel within the device, wherein each biomarker is being tested at two different concentrations (low and high concentrations).

[0316] In Figure 6b, the test lines are configured in a configuration wherein the high concentration test line (215 218) for each biomarker appears closest to the control line and the lowest concentration test line (214, 217) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, test lines 1 and 2 (214 and 215) for BDG are positive, as shown by a solid test line, suggesting a high (above 81 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample and test lines 3 and 4 (217 and 218) are positive, as shown by a solid test line, suggesting a high (0.25 ng / mL or above) concentration of PCT in an (e.g. undiluted) blood sample. This suggests that BDG and PCT are both present in the sample at a high concentration and therefore a bacterial and fungal co-infection is detectable. The clinician may therefore advise that the patient has a fungal and bacterial co-infection present, whereby the fungal and bacterial infections are present to significant levels that would warrant the prescribing of an antifungal and antibiotic agent. The high level of each infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0317] In Figure 6b, there is a solid visual control line (212) indicating that the test has run correctly and so is valid (e.g., the results are valid).

[0318] In Figures 7a-7f, two biomarkers are being tested on a single channel within the device, wherein one biomarker is being tested at a single threshold concentration and one biomarker is being tested at two different concentrations (low and high concentrations).

[0319] In Figure 7b, BDG is tested at a single threshold concentration and PCT is tested at two concentrations (low and high). The test lines for PCT are configured in a configuration wherein the high concentration test line (218) for appears closest to the control line and the lowest concentration test line (217) appears closest to the sample port. In this example, only one test line is present for BDG which is positive as shown by a solid test line, suggesting the presence of BDG at the single threshold concentration (e.g. above 80 pg / mL) in an (e.g. undiluted) blood sample. Test line 2 for BDG is not included within this arrangement device. Test lines 3 and 4 (217 and 218) for PCT are positive, as shown by a solid test line. This suggests a high (0.25 ng / mL or above) concentration of PCT in an (e.g. undiluted) blood sample. This suggests that BDG is present at a desired threshold level and PCT is present at a high concentration. Therefore, a bacterial and fungal infection are detectable in the sample. The clinician may therefore advise that the patient has a fungal and bacterial co-infection wherein the fungal infection is present to a level that an antifungal agent may be prescribed. The patient should be monitored, and clinical judgement used as the infection may be significant enough that it may progress towards sepsis. The high level of bacterial infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0320] In Figure 7c, BDG is tested at a single threshold concentration and PCT is tested at two concentrations (low and high). The test lines for PCT are configured in a configuration wherein the high concentration test line (218) for appears closest to the control line and the lowest concentration test line (217) appears closest to the sample port. In this example, only one test line is present for BDG which is positive as shown by a solid test line, suggesting the presence of BDG at the single threshold concentration (e.g. 80 pg / mL) in an (e.g. undiluted) blood sample. Test line 2 for BDG is not included within this arrangement device. Test line 3 (217) for PCT is positive, as shown by a solid test line, but test line 4 (218) is negative, as there is an absence of any visual solid test line. This suggests a low (0.1-0.24 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. This suggests that BDG is present at a desired threshold level and PCT is present at a low concentration. Therefore, a bacterial and fungal infection may be detectable in the sample. The clinician may therefore advise that the patient has a fungal and bacterial co-infection wherein the fungal infection is present to a level that an antifungal agent may be prescribed. The patient should be monitored, and clinical judgement used as the infection may be significant enough that it may progress towards sepsis. As the bacterial infection is present to a low level the patient should be monitored and clinical judgement used to determine if an antibiotic agent is required at this stage as the level of infection is still low and there is only a low risk of sepsis. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0321] In Figure 7d, BDG is tested at a single threshold concentration and PCT is tested at two concentrations (low and high). The test lines for PCT are configured in a configuration wherein the high concentration test line (218) for appears closest to the control line and the lowest concentration test line (217) appears closest to the sample port. In this example, only one test line is present for BDG which is positive, as shown by a solid test line, suggesting the presence of BDG at the single threshold concentration (e.g. 80 pg / mL) in an (e.g. undiluted) blood sample. Test line 2 for BDG is not included within this arrangement device. Test lines 3 and 4 (217 and 218) for PCT are negative, as shown by the absence of a solid test line. This suggests that PCT is not detected in the sample. This suggests that BDG is present at a desired threshold level but that PCT is not present within the sample. Therefore, a fungal infection is detectable in the sample. The clinician may therefore advise that the patient has a fungal infection wherein the fungal infection is present to a level that an antifungal agent may be prescribed. The patient should be monitored, and clinical judgement used as the infection may be significant enough that it may progress towards sepsis. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor effectiveness of any administered treatment.

[0322] In Figure 7e, BDG is tested at a single threshold concentration and PCT is tested at two concentrations (low and high). The test lines for PCT are configured in a configuration wherein the high concentration test line (218) for appears closest to the control line and the lowest concentration test line (217) appears closest to the sample port. In this example, only one test line is present for BDG which is negative as there is an absence of any visual solid test line. This suggests that BDG is not present in the sample at the single threshold concentration (e.g. 80 pg / mL) in an (e.g. undiluted) blood sample. Test line 2 for BDG is not included within this arrangement device. Test lines 3 and 4 (217 and 218) for PCT are positive, as shown by a solid test line. This suggests a high (0.25 ng / mL or above) concentration of PCT in an (e.g. undiluted) blood sample. This suggests that BDG is not present at a desired threshold level and PCT is present at a high concentration. Therefore, a bacterial infection is detectable in the sample. The clinician may therefore advise that the patient has a bacterial infection present, whereby the bacterial infection is present to a significant level that would warrant the prescribing of an antibiotic agent. The high level of infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0323] Figure 7f shows the same results as Figure 7c, but the two test lines of PCT (low / high) in Figure 7f are configured the opposite way round compared to the test lines of Figure 7c. In Figure 7f, the high concentration line (218) appears closest to the sample port and the low concentration line (217) appears closest to the control line.

[0324] In Figures 7b-7f, there is a solid visual control line (212) indicating that the test has run correctly and so is valid (e.g., the results are valid).

[0325] Example 3

[0326] Figures 8a-15c show example embodiments of the current invention in the form of a multi-channel lateral flow device (201). In these embodiments, the tests have been separated so each biomarker is tested on a different test channel, or some biomarkers are tested in combination with other biomarkers on one channel and further biomarkers are tested on a separate test channel. This may be desirable in order to increase the sensitivity of one of the tests, for example, by using different sample concentrations, reagents, buffers, diluents etc. In particular, certain biomarker tests may be separated out into different channels. This may be desirable in order to increase or decrease the sensitivity of one of the tests, for example, by using different desensitising agents, sensitising agents etc. or to reduce interferences, such as non-specific binding or competition effects.

[0327] The test device (201) is configured as a multi-channel lateral flow device for detecting the presence of biomarkers in a liquid sample. The device comprises a permeable material in the form of porous pads defining: a first portion comprising two separate sample pads accessible via either a single sample port (202) or multiple sample ports (202a, 202b, 202c, 202d) and two separate conjugate pads (not shown) collectively configured for providing a first site for application of the liquid sample and for respective conjugates movably supported therein. The conjugates comprise binders for respective biomarkers coupled to a coloured particle (which may be the different respective colours); and, a second portion comprising two separate test zone pads visible via respective test channels, each defining a respective part of a collective test zone which is configured relative to the two separate conjugate pads of the first portion so as to permit capillary flow communication therebetween. The second portion provides a plurality of separate second sites (114-125, 214-224) each of which is spaced from the first site for visually determining the presence of the coloured particle associated with a respective one of the biomarkers. Each one of the second sites (114-125, 214-224) comprises a binder immobilized therein which binds to a respective one of the biomarkers at a single concentration or at two or more different concentrations of the biomarkers.

[0328] The multichannel lateral flow device (201) essentially operates in the same way as the previous embodiment (101) and uses the same general construction materials. This embodiment may have one sample port (202) which is in direct contact and fluid communication with the respective conjugate pads and test channels, along which the sample picks up the antibody conjugates and becomes capture by their respective immobilised capture antibodies at the relevant test line. Alternatively, the multi-channel lateral flow device may have two, three or four separate sample ports (202a, 202b, 202c, 202d) which are in direct contact and fluid communication with their respective separate conjugate pads and, test channels along which the sample picks up the antibody conjugates and become captured by their respective immobilised capture antibodies at the relevant test line (114-125, 214-224). A separate control line (112a-112d) is present on each test channel) to indicate the validity of each test channels performance.

[0329] In Figures 8a and 8b, two biomarkers are being tested on a multichannel lateral flow device, wherein one biomarker is tested on a first test channel and the second biomarker is tested on a second test channel. Each biomarker is being tested at three different concentrations (low, medium and high concentrations) within each test channel.

[0330] In Figures 8a-10c, test lines 1-3 (114-116) are specific to capturing the BDG-conjugate complex only, where test line 1 (114) has specific immobilised capture antibodies ready to capture BDG-conjugate complexes corresponding to a concentration of 60-80 pg / mL (i.e. a low concentration) in an (e.g. undiluted) blood sample, test line 2 (115) has specific immobilised capture antibodies capture BDG- conjugate complexes corresponding to a concentration of 81-500 pg / mL (i.e. a medium concentration) in an (e.g. undiluted) blood sample and test line 3 (116) has specific immobilised capture antibodies ready to capture BDG-conjugate complexes corresponding to a concentration of above 501 pg / mL (i.e. a high concentration) in an (undiluted) blood sample.

[0331] In Figures 8a-10c, test lines 4-6 (117-119) are specific to capturing the PCT-conjugate complex only, wherein test line 4 (117) has specific immobilised capture antibodies ready to capture PCT-conjugate complexes corresponding to a concentration of 0.1-0.24 ng / mL (i.e. a low concentration) in an (e.g. undiluted) blood sample, test line 5 (118) has specific immobilised capture antibodies capture PCT- conjugate complexes corresponding to a concentration of 0.25-0.49 ng / mL (i.e. a medium concentration) in an (e.g. undiluted) blood sample and test line 6 (1 19) has capture PCT-conjugate complexes corresponding to a concentration of above 0.5 ng / mL (i.e. a high concentration) in an (e.g. undiluted) blood sample.

[0332] In Figures 8a-10c, test lines 7-9 (120-122) in this example will be specific to capturing the MxA-conjugate complex only, wherein test line 7 (120) has specific immobilised capture antibodies ready to capture MxA- conjugate complexes corresponding to a concentration of 40-60 ng / mL (i.e. a low concentration) in an (e.g. undiluted) blood sample, test line 8 (121) has specific immobilised capture antibodies capture MxA- conjugate complexes corresponding to a concentration of 80-100 ng / mL (i.e. a medium concentration) in an (e.g. undiluted) blood sample and test line 9 (122) has specific immobilised capture antibodies ready to capture MxA-conjugate complexes corresponding to a concentration of above 101 ng / mL (i.e. a high concentration) in an (e.g. undiluted) blood sample.

[0333] In Figures 8a-10c, test lines 10-12 (123-125) are be specific to capturing the CRP-conjugate complex only, wherein test line 10 (123) has specific immobilised capture antibodies ready to capture CRP- conjugate complexes corresponding to a concentration of 1 .0-2.0 mg / dL (i.e. a low concentration) in an (undiluted) blood sample, test line 11 (124) has specific immobilised capture antibodies capture CRP- conjugate complexes corresponding to a concentration of 2.1-9.9 mg / dL (i.e. a medium concentration) in an (undiluted) blood sample and test line 12 (125) has specific immobilised capture antibodies ready to capture CRP-conjugate complexes corresponding to a concentration of above 10.0 mg / dL (i.e. a high concentration) in an (undiluted) blood sample.

[0334] In Figure 8a, the multichannel test device (201) has two sample test channels and two sample ports (202a and 202b). The test lines are configured in a configuration wherein the high concentration test line (116, 119) for each biomarker appears closest to the control line and the lowest concentration test line (114, 117) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, test lines 1-3 (114-116) detect BDG in the first test channel of the device. Test lines 1-3 are positive, as shown by a solid test line, suggesting a high (above 501 pg / ml) concentration of BDG in an (e.g. undiluted) blood sample. Test lines 4-6 (117-119) detect PCT in the second test channel of the device. Test lines 4-6 are positive, as shown by a solid test line, suggesting a high (above 0.5 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. This suggests that BDG and PCT are both present in the sample at a high concentration and therefore a bacterial and fungal co-infection is present. The clinician may therefore advise that the patient has a fungal and bacterial co-infection present, whereby the fungal and bacterial infections are present to significant levels that would warrant the prescribing of an antifungal and an antibiotic agent. The high level of each infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0335] Figure 8b shows the same results as Figure 8a but the multichannel test device (201) has a single sample port (202).

[0336] In Figures 8a and 8b, there is a solid visual control line (112a & 112b) on each sample test channel indicating that the test has run correctly and so is valid (e.g., the results are valid).

[0337] In Figures 9a-9c, three biomarkers are being tested on a multichannel lateral flow device, wherein one biomarker is tested on a first test channel, a second biomarker is tested on a second test channel and a third biomarker is tested on a third channel. Each biomarker is being tested at three different concentrations (low, medium and high concentrations) within each test channel.

[0338] In Figure 9a, the multichannel test device (201) has three sample test channels and three sample ports (202a, 202b and 202c). The test lines are configured in a configuration wherein the high concentration test line (116, 119, 122) for each biomarker appears closest to the control line and the lowest concentration test line (114, 117, 120) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, test lines 1-3 (114-116) detect BDG in the first test channel of the device. Test lines 1-3 are positive, as shown by a solid test line, suggesting a high (above 501 pg / ml) concentration of BDG in an (e.g. undiluted) blood sample. Test lines 4-6 (117-119) detect PCT in the second test channel of the device. Test lines 4-6 are positive, as shown by a solid test line, suggesting a high (above 0.5 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test lines 7-9 (120- 122) detect MxA in the third channel of the device. Test lines 7-9 are positive, suggesting a high (above 101 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. This suggests that BDG, MxA and PCT are all present in the sample at a high concentration and therefore a bacterial, viral and fungal coinfection is present. The clinician may therefore advise that the patient has a fungal, bacterial and viral coinfection present, whereby the fungal, bacterial and viral infections are present to significant levels that would warrant the prescribing of an antifungal, antibiotic and an antiviral agent. The high level of each infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0339] Figure 9b shows the same results as Figure 9a but the multichannel test device (201) has a single sample port (202).

[0340] In Figure 9c, the multichannel test device (201) has three sample test channels and a single sample port (202). The test lines are configured in a configuration wherein the high concentration test line (116, 119, 122) for each biomarker appears closest to the sample port and the lowest concentration test line (114, 117, 120) for each biomarker appears closest to the control line for each sub-set of test lines. In this example, test lines 1-3 (114-116) detect BDG in the first test channel of the device. Test lines 1 and 2 (114 and 115) are positive, as shown by a solid test line, but test line 3 (116) is negative in the absence of a solid test line. This suggests that there is a medium (81-500 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample. Test lines 4-6 (117-119) detect PCT in the second test channel of the device. Test lines 4-6 are positive, as shown by a solid test line, suggesting a high (above 0.5 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test lines 7-9 (120-122) detect MxA in the third channel of the device. Test line 7 (120) is positive, as shown by a solid test line, but test lines 8 and 9 (121 and 122) are negative in the absence of a solid test line. This suggests a low (40-60 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. This suggests that BDG is present in the sample at a medium concentration, PCT is present in the sample at a high concentration and MxA is present in the sample at a low concentration. The clinician may therefore advise that the patient has a fungal, bacterial and viral co-infection present, whereby the fungal and bacterial infections are present to significant levels that would warrant the prescribing of an antifungal and an antibiotic agent. The level of each infection suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. As the viral infection is present to a low level the patient should be monitored and clinical judgement used to determine if antiviral agents are required at this stage as the level of infection is still low. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0341] In Figures 9a-9c, there is a solid visual control line (112a, 112b and 12c) on each sample test channel indicating that the test has run correctly and so is valid (e.g., the results are valid).

[0342] In Figures 10a-10c, four biomarkers are being tested on a multichannel lateral flow device, wherein one biomarker is tested on a first test channel, a second biomarker is tested on a second test channel, a third biomarker is tested on a third channel and a fourth biomarker is tested on a fourth test channel. Each biomarker is being tested at three different concentrations (low, medium and high concentrations) within each test channel.

[0343] In Figure 10a, the multichannel test device (201) has four sample test channels and four sample ports (202a, 202b, 202c and 202d). The test lines are configured in a configuration wherein the high concentration test line (116, 119, 122, 125) for each biomarker appears closest to the control line and the lowest concentration test line (114, 1 17, 120, 123) for each biomarker appears closest to the sample port for each sub-set of test lines. In this example, test lines 1-3 (114-116) detect BDG in the first test channel of the device. Test lines 1-3 (114-116) are positive, as shown by a solid test line, suggesting a high (above 501 pg / ml) concentration of BDG in an (e.g. undiluted) blood sample. Test lines 4-6 (117-119) detect PCT in the second test channel of the device. Test lines 4-6 are positive, as shown by a solid test line, suggesting a high (above 0.5 ng / mL) concentration of PCT in an (e.g. undiluted) blood sample. Test lines 7-9 (120-122) detect MxA in the third channel of the device. Test lines 7-9 (120-122) are positive, suggesting a high (above 101 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. Test lines 10-12 (123-125) are positive, as shown by a solid test line, suggesting a high (above 10.0 mg / dL) concentration of CRP in an (e.g. undiluted) blood sample. This suggests that BDG, PCT, MxA and CRP are all present in the sample at a high concentration and therefore a bacterial, fungal and viral infection is detectable along with high levels of inflammation. The clinician may therefore advise that the patient has a fungal, bacterial and viral co-infection present, whereby the infections are present to a significant level that would warrant the prescribing of suitable antimicrobial agents (antifungals, antibiotics and antivirals). The high level of inflammation present suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0344] Figure 10b shows the same results as Figure 10a but the multichannel test device (201) has a single sample port (202).

[0345] In Figure 10c, the multichannel test device (201) has four sample test channels and a single sample port (202). The test lines are configured in a configuration wherein the high concentration test line (116, 119, 122, 125) for each biomarker appears closest to the sample port and the lowest concentration test line (114, 117, 120, 123) for each biomarker appears closest to the control line for each sub-set of test lines. In this example, test lines 1-3 (114-116) detect BDG in the first test channel of the device. Test lines 1 and 2 (114 and 115) are positive, as shown by a solid test line, but test line 3 (116) is negative in the absence of a solid test line. This suggests that there is a medium (81-500 pg / mL) concentration of BDG in an (e.g. undiluted) blood sample. Test lines 4-6 (117-119) detect PCT in the second test channel of the device. Test lines 4-6 are negative in the absence of a solid test line, suggesting that PCT is not present in the sample. Test lines 7-9 (120-122) detect MxA in the third channel of the device. Test line 7 (120) is positive, as shown by a solid test line, but test lines 8 and 9 (121 and 122) are negative in the absence of a solid test line. This suggests a low (40-60 ng / mL) concentration of MxA in an (e.g. undiluted) blood sample. Test lines 10-12 (123-125) are positive, as shown by a solid test line, suggesting a high (above 10.0 mg / dL) concentration of CRP in an (e.g. undiluted) blood sample. This suggests that CRP is present at a high concentration, BDG is present in the sample at a medium concentration, PCT is not present in the sample and MxA is present in the sample at a low concentration. The clinician may therefore advise that the patient has a fungal and viral co-infection present, whereby the fungal infection is present to a significant level that would warrant the prescribing of an antifungal agent. As the viral infection is present to a low level the patient should be monitored and clinical judgement used to determine if an antiviral agent is required at this stage as the level of infection is still low. The high level of inflammation suggests that if not already in hospital, the patient should be admitted for treatment and monitored as an urgent emergency sepsis case. The clinician may choose to monitor and retest the patient again after a defined period of time, to determine if the patient’s condition has changed and to monitor the effectiveness of any administered treatment.

[0346] In Figures 10a-10c, there is a solid visual control line (112a, 112b, 112c and 1 12d) on each sample test channel indicating that the test has run correctly and so is valid (e.g., the results are valid).

[0347] Example 4

[0348] In Example 4, the lateral flow device has the same configuration as the lateral flow device used in Exam...

Claims

Claims:1 . A test device for detecting the presence of biomarkers in a liquid sample, the device comprising: a permeable material defining: a first portion providing a first site for application of the liquid sample and for conjugates movably supported therein wherein each conjugate comprises a binder for a respective biomarker coupled to a respective detector reagent; and, a second portion configured relative to the first portion so as to permit capillary flow communication therebetween, and said second portion providing a second site spaced from the first site for visually determining the presence of the respective detector reagent, and comprising binders immobilized therein each of which binds to a respective said biomarker; wherein the first portion comprises for a first said biomarker, a first said moveably supported conjugate comprising a first binder for a fungal, viral or bacterial biomarker substance coupled to a first respective detector reagent and, for at least one further said biomarker, at least one further said moveably supported conjugate comprising at least one further binder coupled to at least one further detector reagent, wherein the at least one further biomarker comprises a respective biomarker substance selected from a fungal, viral or bacterial biomarker substance, wherein the first biomarker and the at least one further biomarker are different biomarkers, and wherein the second site is configured to visually determine the presence of at least one of the respective detector reagents at two or more different concentrations of at least one of the biomarkers.

2. The test device according to claim 1 , wherein:(i) the first portion comprises a first binder for fungal biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal biomarker substance at two or more different concentrations of the fungal biomarker substance, or,(ii) the first portion comprises a first binder for a fungal biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the bacterial biomarker substance at two or more different concentrations of the bacterial biomarker substance, or,(iii) wherein the first portion comprises a first binder for a fungal biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal and bacterial biomarker substance at two or more different concentrations of the fungal and bacterial biomarker substance respectively, or,(iv) the first portion comprises a first binder for viral biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the viral biomarker substance at two or more different concentrations of the viral biomarker substance, or,(v) the first portion comprises a first binder for a viral biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configuredto visually determine the presence of the bacterial biomarker substance at two or more different concentrations of the bacterial biomarker substance, or,(vi) wherein the first portion comprises a first binder for a viral biomarker substance and at least one further binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the viral and bacterial biomarker substance at two or more different concentrations of the viral and bacterial biomarker substance respectively,(vii) the first portion comprises a first binder for a fungal biomarker substance and at least one further binder for a viral biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal biomarker substance at two or more different concentrations of the fungal biomarker substance, or,(viii) the first portion comprises a first binder for a fungal biomarker substance and at least one further binder for a viral biomarker substance, and wherein the second site is configured to visually determine the presence of the viral biomarker substance at two or more different concentrations of the viral biomarker substance, or,(ix) the first portion comprises a first binder for a fungal biomarker substance and at least one further binder for a viral biomarker substance, and wherein the second site is configured to visually determine the presence of the viral biomarker substance and the fungal biomarker substance at two or more different concentrations of the fungal and viral biomarker substance respectively.

3. The test device according to claim 1 , wherein(i) the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder binds for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal biomarker substance at two or more different concentrations of the fungal biomarker substance, or,(ii) the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the bacterial biomarker substance at two or more different concentrations of the bacterial biomarker substance, or,(iii) wherein the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the viral biomarker substance at two or more different concentrations of the viral biomarker substance, or,(iv) wherein the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal and bacterialbiomarker substance at two or more different concentrations of the fungal and bacterial biomarker substance respectively, or(v) wherein the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal and viral biomarker substance at two or more different concentrations of the fungal and viral biomarker substance respectively, or(vi) wherein the device detects the presence of three biomarkers, wherein the first portion comprises a first binder for fungal biomarker substance, a second binder for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the bacterial and viral biomarker substance at two or more different concentrations of the bacterial and viral biomarker substance respectively, or(vii) wherein the device detects the presence of three biomarkers, wherein the first portion comprises a binder for a fungal biomarker substance, a second binder binds for a viral biomarker substance and a third binder for a bacterial biomarker substance, and wherein the second site is configured to visually determine the presence of the fungal, bacterial and viral biomarker substance at two or more different concentrations of the fungal, bacterial and viral biomarker substance respectively.

4. The test device according to any one of claims 1 -3, wherein the first portion further comprises a binder for a non-specific inflammatory biomarker substance and wherein the second site is configured to visually determine the presence of the non-specific inflammatory biomarker at one concentration or at two or more different concentrations of the non-specific inflammatory biomarker substance; optionally wherein the non-specific inflammatory biomarker substance comprises CRP.

5. The test device according to any one of claims 1 -4, wherein the fungal biomarker substance comprises Beta-D-glucan (BDG).

6. The test device according to any one of claims 1 -4 wherein the viral biomarker substance comprises Myxovirus resistance protein A (MxA).

7. The test device according to any one of claims 1 -4, wherein the bacterial biomarker substance comprises Procalcitonin (PCT).

8. The test device according to any one of claims 1-7, wherein the second site comprises two or more test lines comprising binders immobilised therein which bind to at least one respective biomarker, wherein each test line provides a visual determination of the presence of at least one biomarker at two or more different concentrations of said biomarker, optionally, wherein the first test line provides a visual determination of the presence of said at least one biomarker at a firstconcentration and the second test line provides a visual determination of the presence of said at least one biomarker at a second concentration, optionally, wherein the first test line provides a visual determination of the presence of said at least one biomarker at a low concentration and the second test line provides a visual determination of the presence of said at least one biomarker at a high concentration.

9. The test device according to any one of claims 1-8, wherein the first portion comprises a first binder for a fungal biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal biomarker in the sample, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample.

10. The test device according to any one of claims 1-8, wherein the first portion comprises a first binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL or below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above or about 0.50 ng / mL or above in a blood sample.11 . The test device according to any one of claims 1 -8, wherein the first portion comprises a first binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above in a blood sample.

12. The test device according to any one of claims 1-8, wherein the first portion comprises a first binder for a bacterial biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.50 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.50 ng / mL or above in a blood sample.

13. The test device according to any one of claims 1-8, wherein the first portion comprises a first binder for a viral biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the viral biomarker substance in the sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample.

14. The test device according to any one of claims 1-13, wherein the first portion further comprises a binder for a non-specific inflammatory biomarker substance, and wherein the second site further comprises two or more test lines for visually determining the presence of the non-specific inflammatory biomarker substance in the sample, wherein the non-specific inflammatory biomarker substance comprises CRP and wherein the first test line provides a visual determination of the presence of CRP corresponding to a concentration of below about 9.9 mg / dL in a blood sample and the second test line provides a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

15. The test device according to claims 1-8, wherein the second site comprises three test lines comprising binders immobilised therein which bind to at least one respective biomarker, wherein each test line provides a visual determination of the presence of said biomarker at three or more different concentrations, wherein the first test line provides a visual determination of the presence of said at least one biomarker at a first concentration, the second test line provides a visual determination of the presence of said at least one biomarker at a second concentration, and the third test line provides a visual determination of the presence of said at least one biomarker at a third concentration, optionally, wherein the first test line provides a visual determination of the presence of said at least one biomarker at a low concentration, the second test line provides a visual determination of the presence of said at least one biomarker at a medium concentration, and the third test line provides a visual determination of the presence of said at least one biomarker at a high concentration.

16. The test device according to claim 15, wherein the first portion comprises a first binder for a fungal biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the fungal biomarker substance in the sample, wherein the fungal biomarker comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample, the second test line provides a visual determination of BDG corresponding to a concentration of about 81-500 pg / mL in a blood sample and the third test line provides a visual determination of BDG corresponding to a concentration of above about 500 pg / mL in a blood sample.

17. The test device according to claim 15, wherein the first portion comprises a first binder for a viral biomarker substance, and wherein the second site comprises three or more test lines for visuallydetermining the presence of the viral biomarker substance in the sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample, the second test line provides a visual determination of the presence of MxA corresponding to a concentration of about 61-100 ng / mL in a blood sample and the third test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 100 ng / mL in a blood sample.

18. The test device according to claim 15, wherein the first portion comprises a first binder for a bacterial biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the bacterial biomarker substance in the sample, wherein the bacterial biomarker substance comprises PCT, wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample, the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25-0.49 ng / mL in a blood sample and the third test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.5 ng / mL or above in a blood sample.

19. The test device according to any one of claims 1-18, wherein the first portion further comprises a binder for a non-specific inflammatory biomarker substance, and wherein the second site further comprises three or more test lines for visually determining the presence of the non-specific inflammatory biomarker substance in the sample, wherein the non-specific inflammatory biomarker substance comprises CRP and wherein the first test line provides a visual determination of the presence of CRP corresponding to a concentration of below about 2.0 mg / dL in a blood sample, the second test line provides a visual determination of the presence of CRP corresponding to a concentration of about 2.1-9.9 mg / dL in a blood sample and the third test line provides a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

20. The test device according to any one of claims 1-8, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance, a third binder for a bacterial biomarker substance and a fourth binder for a non-specific inflammatory biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal, bacterial, viral and non-specific inflammatory biomarker substance in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presenceof MxA corresponding to a concentration of above about 60 ng / mL in a blood sample, wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25 ng / mL or above in a blood sample, and wherein the non-specific inflammatory marker comprises C-reactive protein (CRP), and wherein the first test line provides a visual determination of the presence of CRP corresponding to a concentration of below about 9.9 mg / dL in a blood sample and the second test line provides a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.21 . The test device according to any one of claims 1 -8, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance, a third binder for a bacterial biomarker substance and a fourth binder for a non-specific inflammatory biomarker substance, and wherein the second site comprises two or more test lines for visually determining the presence of the fungal, bacterial, viral and non-specific inflammatory biomarker substance in the sample each at two or more different concentrations, wherein the fungal biomarker substance comprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample and the second test line provides a visual determination of BDG corresponding to a concentration of above about 80 pg / mL in a blood sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample and the second test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 60 ng / mL in a blood sample, wherein the bacterial biomarker substance comprises PCT, and wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.5 ng / mL in a blood sample and the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.5 ng / mL or above in a blood sample, and wherein the non-specific inflammatory marker comprises C-reactive protein (CRP), and wherein the first test line provides a visual determination of the presence of CRP corresponding to a concentration of below about 9.9 mg / dL in a blood sample and the second test line provides a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

22. The test device for any one claims 1 -8, wherein the first portion comprises a first binder for a fungal biomarker substance, a second binder for a viral biomarker substance, a third binder for a bacterial biomarker substance and a fourth binder for a non-specific inflammatory biomarker substance, and wherein the second site comprises three or more test lines for visually determining the presence of the fungal, viral, bacterial and non-inflammatory biomarker substances in the sample each at three different concentrations, wherein the fungal biomarkercomprises BDG and wherein the first test line provides a visual determination of the presence of BDG corresponding to a concentration of about 80 pg / mL or below in a blood sample, the second test line provides a visual determination of BDG corresponding to a concentration of about 81- 500 pg / mL in a blood sample and the third test line provides a visual determination of BDG corresponding to a concentration of above about 500 pg / mL in a blood sample, wherein the viral biomarker substance comprises MxA and wherein the first test line provides a visual determination of the presence of MxA corresponding to a concentration of about 60 ng / mL or below in a blood sample, the second test line provides a visual determination of the presence of MxA corresponding to a concentration of about 61-100 ng / mL in a blood sample and the third test line provides a visual determination of the presence of MxA corresponding to a concentration of above about 100 ng / mL in a blood sample, wherein the bacterial biomarker substance comprises PCT, wherein the first test line provides a visual determination of the presence of PCT corresponding to a concentration of below about 0.25 ng / mL in a blood sample, the second test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.25-0.49 ng / mL in a blood sample and the third test line provides a visual determination of the presence of PCT corresponding to a concentration of about 0.5 ng / mL or above in a blood sample, and wherein the non-specific inflammatory biomarker substance comprises CRP, wherein the first test line provides a visual determination of the presence of CRP corresponding to a concentration of below about 2.0 mg / dL in a blood sample, the second test line provides a visual determination of the presence of CRP corresponding to a concentration of about 2.1 -9.9 mg / dL in a blood sample and the third test line provides a visual determination of the presence of CRP corresponding to a concentration of about 10.0 mg / dL or above in a blood sample.

23. The test device according to any one of the preceding claims wherein the liquid sample comprises a blood sample.

24. A test device according to any preceding claim wherein the permeable material is planar and the second site is laterally spaced from the first site along the planar permeable material such that the test device provides a lateral flow test device.

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