Fluid transfer device having an integrated flow-based assay and method of using the same for sepsis identification

Abstract

A system for the early detection and treatment of sepsis, comprising a fluid transfer device having an inlet and an outlet. The inlet is configured to receive a flow of body fluid. A flow-based assay device is configured to be coupled to the fluid transfer device. A portion of the flow-based assay device engages the outlet to enable a portion of a first volume of body fluid to be transferred to the flow-based assay device. The flow-based assay device is configured to detect at least one sepsis-related biomarker. A processing circuit receives data from the flow-based assay device, applies a computational model to the received data, generates a sepsis probability score based on an output of the applied computational model, and alerts a healthcare provider to initiate a corresponding treatment when the sepsis probability score exceeds a sepsis-related threshold.

Description

Technical Field

[0001] (Cross - Reference to Related Applications) This application claims the benefit and priority of U.S. Provisional Patent Application No. 63 / 345,106, entitled "Fluid Transfer Devices with Integrated Flow - Based Assay and Methods of Using the Same for Identifying Sepsis," filed on May 24, 2022, the disclosure of which is incorporated herein by reference in its entirety.

[0002] This application is related to International Patent Application No. PCT / US2020 / 064600, entitled "Fluid Transfer Devices with Integrated Flow - Based Assay and Methods of Using the Same," filed on December 11, 2020, and U.S. Patent Application No. 17 / 119,732, entitled "Fluid Transfer Devices with Integrated Flow - Based Assay and Methods of Using the Same," filed on December 11, 2020. Each of these claims the benefit and priority of U.S. Provisional Patent Application No. 62 / 946,680, entitled "Fluid Transfer Devices with Integrated Flow - Based Assay and Methods of Using the Same," filed on December 11, 2019, and the disclosure of each of these is incorporated herein by reference in its entirety.

[0003] The embodiments described herein generally relate to the acquisition of body fluid samples and point - of - care diagnostic tests, and more particularly, to body fluid transfer devices having an integrated flow - based assay system for, e.g., initial point - of - care diagnostic tests for sepsis.

Background Art

[0004] Medical practitioners routinely perform a wide variety of diagnostic tests on patients using parenterally obtained body fluids for various types of microorganisms and others. In some instances, such as certain severe patient situations (e.g., sepsis), effective treatment is time-dependent, and treatment delays can potentially lead to an increased risk of morbidity and / or mortality. For example, sepsis, which generally results from a bacterial infection (or less commonly a fungal or viral infection), is an abnormal systemic reaction to what can be a normal infection and is likely to represent a pattern of response by the immune system to injury. After an excessive inflammatory response, there is generally multiple organ dysfunction and a subsequent immunosuppressed phase in which the patient is susceptible to nosocomial infections. Septic patients typically present with malaise, fever, chills, and leukocytosis, which can prompt physicians to evaluate such patients for the presence of bacteria in the bloodstream, typically via a bacterial culture test.

[0005] As bacterial culture testing and other advanced diagnostic techniques evolve and improve, the speed, accuracy (both sensitivity and specificity), and value of the information that can be provided to clinicians continue to increase. Examples of such diagnostic techniques can include, for example, microbial detection, molecular diagnostics, gene sequencing (such as deoxyribonucleic acid (DNA), ribonucleic acid (RNA), next-generation sequencing (NGS), etc.), biomarker identification, and the like. However, due to the time scales associated with these diagnostic techniques, most treatment decisions need to be made before blood culture results can be returned and are made based on physiological judgments. Furthermore, some known culture methods and / or other diagnostic techniques can tend to be contaminated, thereby resulting in inaccurate, distorted, adulterated, false-positive, false-negative, and / or in other cases results that do not represent the patient's actual condition (or in vivo situation). Consequently, these results can potentially lead to incorrect, inaccurate, confused, uncertain, unreliable, and / or in other cases undesirable clinical decisions. Additionally, in some instances, contamination can be caused by the presence of biological substances containing cells external to the intended sample source and / or other external contaminants inadvertently included in the body fluid sample being analyzed.

[0006] Even if a clean or pure body fluid is provided, tests from such diagnostic techniques can take from 6 hours to over 5 days to obtain results, are often performed using systems that require highly trained personnel, and / or often use culture protocols specially adjusted for the identification of various bacterial species. Furthermore, sepsis can progress rapidly to multiple organ failure and / or death, and physicians may have to prescribe therapeutic agents (e.g., antibiotics) before receiving the results of diagnostic tests. In fact, current guidelines for adults at risk of septic shock or highly likely to have sepsis recommend the immediate administration of antibacterial agents, ideally within 1 hour of recognition. When septic shock or sepsis is uncertain, the guidelines recommend continuous patient evaluation for indicators that suggest sepsis or indicators that suggest no sepsis, which is a challenging task for a rapidly progressing disease. Thus, such culture methods and / or diagnostic techniques are not suitable for the rapid diagnosis and / or efficient screening that may be required to treat certain rapidly progressing diseases.

[0007] Accordingly, there is a need for rapid testing of body fluids, such as point-of-care diagnostic tests that use lateral flow assays or other rapid diagnostic techniques for the detection and / or prediction of sepsis. In addition, it is necessary to integrate a rapid test (e.g., a lateral flow assay) into a device that can be used to obtain additional body fluid samples from a patient, e.g., a device configured to obtain a body fluid sample with reduced contamination. SUMMARY OF THE INVENTION

[0008] According to one embodiment, the present disclosure is a system for early detection and treatment of sepsis, comprising a fluid transfer device having an inlet and an outlet, the inlet being configured to receive a flow of body fluid from a body fluid source, and at least one flow-based assay device configured to be coupled to the fluid transfer device, a part of the at least one flow-based assay device engaging the outlet when coupled to the fluid transfer device to enable a portion of a first volume of body fluid to be transferred from the fluid transfer device to the at least one flow-based assay device, and the at least one flow-based assay device being configured to detect at least one sepsis-related biomarker.

[0009] According to one embodiment, the present disclosure is a system for the early detection and treatment of sepsis, comprising a fluid transfer device having an inlet and an outlet, the inlet being configured to receive a flow of body fluid from a body fluid source, at least one flow-based assay device configured to be coupled to the fluid transfer device, a portion of the at least one flow-based assay device being configured to engage the outlet when coupled to the fluid transfer device to enable a portion of a first volume of body fluid to be transferred from the fluid transfer device to the at least one flow-based assay device, the at least one flow-based assay device being configured to detect at least one sepsis-related biomarker, a processing circuit, the processing circuit being configured to receive data corresponding to at least one sepsis-related biomarker from the at least one flow-based assay device, apply a computational model to the received data, generate a sepsis probability score based on the output of the applied computational model, and alert a healthcare provider to initiate a corresponding treatment when the sepsis probability score exceeds a sepsis-related threshold. In one embodiment, the at least one flow-based assay device is one of a sandwich lateral flow assay device or a competitive lateral flow assay device. In one embodiment, the at least one flow-based assay device includes a conjugate element including a labeled bioactive agent configured to bind to at least one sepsis-related biomarker and one or more capture elements configured to immobilize each at least one sepsis-related biomarker and the labeled bioactive agent, and the accumulation of the labeled bioactive agent immobilized along the one or more capture elements is configured to provide a visual indicator associated with the presence of each at least one sepsis-related biomarker in a portion of the first volume of body fluid. In one embodiment, the labeled bioactive agent includes at least one of an antibody, an aptamer, and a protein binder. In one embodiment, the body fluid is blood.In one embodiment, at least one sepsis-related biomarker is one of procalcitonin, lactate, surface antigen classification 64, neutrophil count marker, and interleukin 6. In one embodiment, the neutrophil count marker is one of neutrophil elastase, lactoferrin, myeloperoxidase, and human neutrophil lipocalin. In one embodiment, at least one sepsis-related biomarker is surface antigen classification 64 and the neutrophil count marker. In one embodiment, the computational model is based on a random forest classifier. In one embodiment, the computational model is trained on a reference dataset that includes at least one of point-of-care metrics and historical metrics. In one embodiment, the point-of-care metrics include one or more of lactate, interleukin 6, surface antigen classification 64, procalcitonin, neutrophil count marker, heart rate, blood pressure, white blood cell count, respiratory rate, and body temperature. In one embodiment, the neutrophil count marker is one of neutrophil elastase, lactoferrin, myeloperoxidase, and human neutrophil lipocalin. In one embodiment, the historical metrics can be derived from one or more of medical history data, previous diagnoses, treatment plans and medications, laboratory and test results, immunization details and dates, and medical images.

[0010] According to one embodiment, the present disclosure is a method for the early detection and treatment of sepsis, comprising receiving data related to at least one flow-based assay device corresponding to at least one sepsis-related biomarker, applying a computational model to the received data, generating a sepsis probability score based on the output of the applied computational model, and warning a healthcare provider to initiate a corresponding treatment when the sepsis probability score exceeds a sepsis-related threshold.

[0011] According to one embodiment, the present disclosure is a method for the early detection and treatment of sepsis, comprising placing an inlet of a fluid transfer device in fluid communication with a body fluid source, receiving a body fluid into the fluid transfer device from the inlet, establishing fluid communication between the inlet and the outlet of the fluid transfer device to enable a volume of the body fluid to flow to a sample reservoir in fluid communication with the outlet, conveying a portion of the volume of the body fluid to a sample element of at least one flow-based assay device fluidly connected to the fluid transfer device at least temporarily, and conveying a buffer solution to the sample element of the at least one flow-based assay device.

Brief Description of the Drawings

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DETAILED DESCRIPTION OF THE INVENTION

[0013] Definitions As used in this specification and / or any claims contained herein, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, the term "member" is intended to mean a single member or a combination of members, and the term "material" is intended to mean one or more materials, etc.

[0014] As used herein, when used in connection with a recited value or geometric configuration or relationship, the terms “about,” “approximately,” and / or “substantially” are intended to convey that the so-defined value or characteristic is the value or characteristic as nominally described. In some instances, the terms “about,” “approximately,” and / or “substantially” generally mean within a desired tolerance (e.g., plus or minus 10% of the recited value or characteristic), and / or can be assumed. For example, a value of about 0.01 can include from 0.009 to 0.011, a value of about 0.5 can include from 0.45 to 0.55, a value of about 10 can include from 9 to 11, and a value of about 100 can include from 90 to 110. Similarly, if a surface is nominally parallel, the first surface can be described as being substantially parallel to the second surface. The recited values, structures, and / or relationships may be desirable, but it should be understood that some variation can occur, for example, as a result of manufacturing tolerances or other practical considerations (e.g., pressure or force applied through a portion of a device, conduit, lumen, etc.). Thus, the terms “about,” “approximately,” and / or “substantially” can be used herein to account for such tolerances and / or considerations.

[0015] As used herein, “body fluid” can include any fluid obtained directly or indirectly from a patient's body. For example, “body fluid” includes, but is not limited to, blood, cerebrospinal fluid, urine, bile, lymph fluid, saliva, synovial fluid, serous fluid, pleural fluid, amniotic fluid, mucus, sputum, vitreous humor, air, etc., or any combination thereof.

[0016] As used herein, the terms “proximal” and “distal” refer to the direction of approaching and the direction of moving away, respectively, for a user placing the device in contact with a patient. Thus, for example, the end of the device that first touches the patient's body is the distal end, and the opposite end of the device (e.g., the end of the device being operated by the user) is the proximal end of the device.

[0017] As used herein, the terms “first,” “initial,” and / or “pre-sample” are used interchangeably when used to describe a volume of a body fluid to describe the amount, portion, or volume of the body fluid from which collection, diversion, isolation, testing, etc. is performed prior to obtaining a “sample” volume. The “first,” “initial,” and / or “pre-sample” volume can be a predetermined volume, a defined volume, a desired volume, and / or a given volume of the body fluid. For example, a predetermined and / or desired pre-sample volume of a body fluid such as blood can be one drop of blood, several drops of blood, about 0.1 milliliter (mL), about 0.2 mL, about 0.3 mL, about 0.4 mL, about 0.5 mL, about 0.6 mL, about 0.7 mL, about 0.8 mL, about 0.9 mL, about 1.0 mL, about 2.0 mL, about 3.0 mL, about 4.0 mL, about 5.0 mL, about 6.0 mL, about 7.0 mL, about 8.0 mL, about 9.0 mL, about 10.0 mL, about 20.0 mL, about 50.0 mL of volume, and / or any volume or fraction of a volume therebetween. In other examples, the pre-sample volume can be greater than 50 mL or less than 0.1 mL. In some particular embodiments, the predetermined and / or desired pre-sample volume can be from about 0.1 mL to about 5.0 mL. As another example, the pre-sample volume can be, for example, the total volume of any number of lumens (e.g., the lumen of a needle and / or the main lumen forming at least a part of the flow path from the body fluid source to an initial collection chamber, portion, reservoir, etc.). As yet another example, the pre-sample volume can be, for example, the volume of the body fluid sufficient to perform an initial or pre-sample test such as a rapid diagnostic test using a lateral flow assay and / or any other rapid test device.

[0018] As used herein, the terms "second," "subsequent," and / or "sample," when used to describe a volume of a body fluid, can be used interchangeably to describe the amount, portion, or volume of body fluid collected after the first volume of body fluid, initial volume, and / or pre-sample volume. The "second," "subsequent," and / or "sample" volume can be a random volume of body fluid collected after collecting, diverting, and / or sequestering the pre-sample volume of body fluid, or a predetermined or desired volume. For example, in some instances, the desired sample volume of body fluid can be from about 10 mL to about 60 mL. In other instances, the desired sample volume of body fluid can be less than 10 mL or greater than 60 mL. In yet other instances, for example, the sample volume can be based at least in part on one or more tests, evaluations, analyses, and / or processes performed on the sample volume.

[0019] In some implementations, the second, subsequent, and / or sample volume of body fluid can be used, for example, in one or more sample or diagnostic tests such as a culture test. In some instances, collecting the "sample" volume of body fluid after collecting, sequestering, isolating, and / or testing the "pre-sample" volume of body fluid can reduce the likelihood that the sample volume contains contaminants such as microorganisms present on the skin. Thus, the sample volume of body fluid can be suitable for highly sensitive tests that might otherwise produce inaccurate results due to contamination.

[0020] The embodiments and / or portions thereof described in this specification can be formed or constructed with one or more biocompatible materials. In some embodiments, the biocompatible material can be selected based on one or more properties of the constituent materials, such as, for example, rigidity, toughness, durometer, bioreactivity, etc. Examples of suitable biocompatible materials include metals, glasses, ceramics, or polymers. Examples of suitable metals include pharmaceutical grade stainless steel, gold, titanium, nickel, iron, platinum, tin, chromium, copper, and / or their alloys. The polymer material can be biodegradable or non-biodegradable. Examples of suitable biodegradable polymers include polylactide, polyglycolide, polylactide-co-glycolide (PLGA), polyanhydrides, polyorthoesters, polyether esters, polycaprolactone, polyester amides, poly(butyric acid), poly(valeric acid), polyurethanes, and / or their blends and copolymers. Examples of non-biodegradable polymers include nylon, polyester, polycarbonate, polyacrylate, polysiloxane (silicone), ethylene vinyl acetate and other acyl-substituted cellulose acetate polymers, non-degradable polyurethanes, polystyrene, polyvinyl chloride, polyvinyl fluoride, poly(vinyl imidazole), chlorosulfonated polyolefin, polyethylene oxide, and / or their mixtures and copolymers.

[0021] "Sepsis" refers to the systemic host response to infection and occurs when an existing infection triggers a chain reaction throughout the body. As used herein, sepsis includes all stages of sepsis, including, but not limited to, the onset of sepsis, severe sepsis, septic shock, and multiple organ failure associated with the final stages of sepsis. "Onset of sepsis" refers to the early stage of sepsis, for example, prior to the stage where conventional clinical symptoms are sufficient to support a clinical suspicion of sepsis. Since the methods of the present invention are used to detect sepsis before the time when sepsis is suspected using conventional techniques, the disease status of a patient with early sepsis can only be retrospectively confirmed when the signs of sepsis are more clinically apparent. The methods of the present invention can detect the onset of sepsis regardless of the origin of the infection process.

[0022] Current guidelines for the treatment of possible septic shock or high likelihood of sepsis promote rapid treatment with antibiotics within 1 hour of recognition. Blood cultures, the "gold standard" for the detection of systemic infection, can take up to 2 days to reliably provide a "negative" result. Furthermore, blood cultures rely on the presence of bacteremia. If blood cultures are drawn after the initiation of antibiotic therapy, growth can be suppressed. Due to this delay and uncertainty, healthcare providers often have to rely on clinical symptoms and less predictive laboratory measurements such as fever or white blood cell (WBC) count when trying to document the presence of infection. Newer markers such as C-reactive protein and procalcitonin are being used to enhance diagnostic sensitivity and specificity. Surface antigen classification 64 (CD64, high-affinity Fcγ receptor) has been shown to increase in patients with bacterial infection. Several studies have shown that measurement of surface granulocyte CD64 may be useful for the detection of bacterial infection and sepsis in the workup of patients with systemic inflammatory response syndrome and fever of unknown origin.

[0023] The CD64 index may be a promising test for the detection and monitoring of antibacterial therapy, but this test has a practical limitation in that it currently has to be performed on a flow cytometer. This requires access to both the flow cytometer and an operator. Typically, flow cytometers are located in laboratories that operate only during business hours and on weekdays. Adding tests that need to be available 24 hours a day, 7 days a week would mean significant reconfiguration of these laboratories.

[0024] Considering the guidelines from the Surviving Sepsis Campaign and the limitations outlined above regarding, for example, CD64 measurement, it can be understood that it is difficult to provide all the necessary test data on sepsis-related biomarkers within a suitable time frame. Therefore, a rapid diagnostic approach for such sepsis-related biomarkers is needed.

[0025] Any of the fluid transfer devices described herein can be configured to receive, acquire, and / or transfer a body fluid flow, bolus, volume, and the like. Additionally, any of the fluid transfer devices described herein can include an integrated device for performing one or more rapid diagnostic tests on at least a portion of the body fluid acquired by the fluid transfer device. In some embodiments, the fluid transfer device can be a syringe, transfer adapter, and / or any other device configured to receive a body fluid flow. In some embodiments, the fluid transfer device can be a fluid diversion and / or isolation device configured to receive and isolate an initial volume of body fluid from a subsequent sample volume, such as in a culture test. In such embodiments, the integrated device for rapid diagnostic testing can be configured to receive at least a portion of the initial volume of body fluid or at least a portion of the subsequent sample volume. The integrated device for rapid diagnostic testing can be, for example, a lateral flow assay and / or any other suitable diagnostic test device. The integrated device for rapid diagnostic testing can be used to examine a volume of body fluid and provide at least a qualitative result, which can then be output on or by a device for visual inspection. In other examples, the test device can communicate data associated with the result to an electronic device (e.g., via a wired or wireless network), and the electronic device can then perform any suitable analysis on the data, such as graphically representing at least some of the data (e.g., qualitative or quantitative test results) on a display of the device.

[0026] In some implementations, the rapid diagnostic test device may be included in or integrated with a fluid transfer device (e.g., a sample collection device) and used to provide an initial test result of the acquired body fluid. The initial test result can be supplemented using additional tests of the acquired body fluid, such as a culture test. For example, an integrated rapid diagnostic test device (also referred to herein as a “rapid test device” or “initial test device”) can provide a method for performing a relatively rapid test of a body fluid for the presence of a microorganism (e.g., a gram-positive bacterium, a gram-negative bacterium, a fungus, or a virus) or other type of biological substance (e.g., a specific type of cell, biomarker, protein, antigen, enzyme, blood component, etc.), which can inform a clinician's decision-making regarding treatment strategies. In some implementations, the initial test device can test for bacteria and / or other infections that can lead to and / or otherwise result in sepsis, thereby enabling a clinician to provide rapid treatment such as broad-spectrum antibiotics. Further, the fluid transfer devices described herein can be used to obtain additional sample volumes for use in other more sensitive tests such as culture tests, or molecular polymerase chain reaction (PCR), magnetic resonance and other magnetic analysis platforms, automated microscopy, spatial clone isolation, flow cytometry, whole blood (“culture-free”) specimen analysis (e.g., NGS) and related techniques, morphological kinetic cell analysis, and / or other general, advanced, or evolving techniques such as other techniques used to characterize a patient's specimen and / or to detect, identify, classify, categorize, and / or characterize a particular organism, antibiotic susceptibility, etc.

[0027] In some embodiments, the system includes at least one flow-based assay device and a fluid transfer device. The fluid transfer device includes an inlet configured to be placed in fluid communication with a body fluid source and an outlet configured to be placed in fluid communication with a sample reservoir. The fluid transfer device includes an isolation chamber and a port that selectively communicates with the isolation chamber. The isolation chamber is configured to be placed in fluid communication with the inlet to receive a first volume of body fluid when the fluid transfer device is in a first state. The outlet is configured to be placed in fluid communication with the inlet to receive a second volume of body fluid when the fluid transfer device is in a second state. The at least one flow-based assay device is configured to be coupled to the port to receive a portion of the first volume of body fluid when the fluid transfer device is in a third state. The at least one flow-based assay device is configured to provide an indicator associated with the presence of a target analyte in a portion of the first volume of body fluid.

[0028] In some embodiments, the system includes a fluid transfer device having an inlet configured to receive a flow of body fluid from a body fluid source, an outlet configured to be placed in fluid communication with a sample reservoir, an isolation chamber configured to receive a first volume of body fluid, and a port in at least temporary fluid communication with the isolation chamber. The fluid transfer device is configured to transition between a first state in which the isolation chamber is in fluid communication with the inlet to receive a first volume of body fluid and a second configuration in which the outlet is in fluid communication with the inlet to receive a second volume of body fluid. The port of the isolation chamber enables a flowing gas to flow through the isolation chamber when the isolation chamber receives the first volume of body fluid. The flow-based assay device is configured to be coupled to the fluid transfer device in the second state. A portion of the flow-based assay device is configured to engage the port when coupled to the fluid transfer device to enable a portion of the first volume of body fluid to be transferred from the isolation chamber to the flow-based assay device. The flow-based assay device is configured to provide an indicator associated with the presence of a target analyte in a portion of the initial volume of body fluid.

[0029] In some embodiments, the method includes placing the inlet of the fluid transfer device in fluid communication with a body fluid source and receiving a first volume of body fluid from the inlet into the isolation chamber of the fluid transfer device, wherein a flow controller of the fluid transfer device enables a flow of gas through the flow controller but not a flow of body fluid and vents the isolation chamber during the receiving. After the first volume of body fluid is received in the isolation chamber, the fluid transfer device is transitioned from the first state to the second state. In response to the fluid transfer device being in the second state, fluid communication is established between the inlet and the outlet of the fluid transfer device to enable a second volume of body fluid to flow to a sample reservoir in fluid communication with the outlet. Transferring a portion of the first volume of body fluid from the isolation chamber to a sample element of a flow-based assay device that is at least temporarily fluidly coupled to the isolation chamber, and transferring a buffer solution to the sample element of the flow-based assay device.

[0030] In some embodiments, the system includes a fluid transfer device and a lateral flow assay device. The fluid transfer device includes an inlet configured to be placed in fluid communication with a body fluid source, an outlet configured to be placed in fluid communication with a sample reservoir, and an isolation chamber configured to receive an initial volume of the body fluid. The fluid transfer device is configured to transition between (1) a first state in which the isolation chamber is in fluid communication with the inlet to receive an initial volume of the body fluid, (2) a second state in which the outlet is in fluid communication with the inlet to receive a subsequent flow of the body fluid, and (3) a third state in which the lateral flow assay device is coupled to a port in fluid communication with the isolation chamber. The lateral flow assay device is configured to receive a portion of the initial volume of the body fluid and determine the presence of a target analyte in the initial volume of the body fluid.

[0031] In some embodiments, the system includes a fluid transfer device and at least one lateral flow assay device configured to detect one or more sepsis-related biomarkers. The fluid transfer device includes an inlet configured to be placed in fluid communication with a body fluid source, an outlet configured to be placed in fluid communication with a sample reservoir, and an isolation chamber configured to receive an initial volume of the body fluid. The fluid transfer device is configured to transition between (1) a first state in which the isolation chamber is in fluid communication with the inlet to receive an initial volume of the body fluid, (2) a second state in which the outlet is in fluid communication with the inlet to receive a subsequent flow of the body fluid, and (3) a third state in which at least one lateral flow assay device is coupled to a port in fluid communication with the isolation chamber. The at least one lateral flow assay device is configured to receive a portion of the initial volume of the body fluid and determine the presence of one or more sepsis-related biomarkers in the initial volume of the body fluid. The one or more sepsis-related biomarkers can include a neutrophil count marker and CD64.

[0032] Referring now to the drawings, FIG. 1 is a schematic diagram of a fluid transfer and assay system 100 according to one embodiment. Although various components, elements, features, and / or functions may be described below, it should be understood that they are presented by way of example only and not by way of limitation. Those skilled in the art will understand that the form and / or features of the fluid transfer and assay system 100 can be modified without changing the ability of the fluid transfer and assay system 100 to acquire a body fluid sample and perform the function of providing a rapid diagnostic test method, as described herein.

[0033] The fluid transfer and assay system 100 (also referred to herein as the "system") can include at least a fluid transfer device 105 and a rapid diagnostic test device 170. In some implementations, the system 100 can optionally include at least one electronic device 190 and / or at least one fluid collection device 195.

[0034] The fluid transfer device 105 (also referred to herein as the "transfer device") can be of any suitable shape, size, and / or configuration, as described herein with reference to particular embodiments. In some implementations, the transfer device 105 can be configured to draw a body fluid (e.g., blood) from a patient into the transfer device 105. Additionally, the transfer device 105 can be configured to transfer at least a portion of the drawn body fluid to one or more other devices, reservoirs, containers, vials, machines, tests, assays, etc., such as one or more of the rapid diagnostic test device 170 and / or the optional fluid collection device 195.

[0035] In some embodiments, the transfer device 105 is configured to transfer, direct, and / or divert a particular amount or volume of body fluid into (or through) one or more portions of the transfer device 105 and then transfer such amount or volume into one or more sample reservoirs, containers, bottles, etc., within one or more devices coupled or integrated with the transfer device 105. For example, the transfer device 105 may be configured to transfer a first portion, amount, or volume of body fluid into or through a first portion or isolation portion of the transfer device 105 and then transfer a second portion, amount, or volume (e.g., a subsequent amount) of body fluid into a second portion or sampling portion of the transfer device 105. In some embodiments, the transfer device 105 and / or the isolation portion of the transfer device 105 may be configured to isolate a first amount of body fluid (e.g., within the isolation portion of the transfer device 105) from a subsequent amount of body fluid, as described in more detail herein with reference to particular embodiments. In some implementations, the transfer device 105 may be configured to transfer at least a portion of a first amount of body fluid (e.g., contained within the isolation portion of the transfer device 105) to the rapid diagnostic test device 170 and transfer at least a portion of a second amount of body fluid to one or more of the optional fluid collection devices 195.

[0036] The rapid diagnostic test device 170 (also referred to herein as the "rapid test device" or simply the "test device") can be of any suitable shape, size, and / or configuration, as described herein with reference to particular embodiments. In some embodiments, the rapid test device 170 can be removably coupled to the transfer device 105 or any suitable portion thereof (e.g., an inlet portion, an outlet portion, an isolation portion, a sampling portion, and / or any other suitable portion). In other embodiments, the rapid test device 170 can be integrated into the transfer device 105. For example, the transfer device 105 and the rapid test device 170 can be formed integrally or monolithically and / or integrated in other ways. In still other embodiments, the transfer device 105 can include, and / or can form, ports, adapters, and / or receiving portions to which the rapid test device 170 can be coupled, or ports, adapters, and / or receiving portions into which the rapid test device 170 can be inserted to establish fluid communication therebetween. In some such embodiments, coupling the rapid test device 170 to the transfer device 105 can act to transition one or more flow controllers, valves, partitions, ports, seals, etc. from a closed or sealed state to an open state to enable fluid communication between the transfer device 105 and the test device 170.

[0037] In some embodiments, the rapid test device 170 can be configured to receive a first quantity of body fluid from the transfer device 105 and perform one or more tests, assays, and / or diagnostic methods using the first quantity of body fluid. For example, the rapid test device 170 can be a chromatographic lateral flow immunoassay capable of testing for any suitable analyte, biomarker, protein, molecule, particle, etc., such as sepsis-related biomarkers. A chromatographic lateral flow immunoassay (referred to herein as a "lateral flow assay" or "LFA") is typically a nitrocellulose-based device configured to detect the presence of a target analyte in a sample (e.g., a biological sample and / or a body fluid sample such as blood, urine). Generally, an LFA includes a series of capillary beds, such as porous paper, microstructured or sintered polymers, that can be arranged in a desired position and / or array configuration on a substrate to direct the flow of a sample (e.g., at least a portion of the first quantity of body fluid) along a portion of the LFA.

[0038] The LFA can be used for a wide range of applications that desirably have a relatively fast, easy-to-use, and low-cost method for rapid antigen detection. The LFA is typically performed with little or no sample or reagent preparation, which can enable useful test results in just a few minutes (or, if more sensitive test results are desired, a longer time). Further, in some embodiments, the LFA can be configured to test for analytes and / or biomarkers produced by the human body in response to in vivo conditions (e.g., an infectious disease such as sepsis), which can mean that such an LFA has a relatively low sensitivity to contaminants (e.g., microorganisms present on the skin) that may be contained in the first quantity of body fluid withdrawn from a patient via the transfer device 105.

[0039] Typically, two types of LFAs are used depending on the size and / or number of binding sites on the target analyte. Specifically, competitive LFAs are generally used when examining smaller analytes, and sandwich LFAs are generally used when examining larger analytes. In context, home pregnancy tests are well-known sandwich lateral flow assays. In some examples, it may be desirable to use a sandwich LFA to examine antigens, analytes, and / or biomarkers associated with sepsis and / or other infectious conditions in a sample of body fluid such as blood. It should be understood that the embodiments described herein include and / or implement sandwich LFAs, but the embodiments are not limited thereto. For example, any of the embodiments described herein can use and / or implement competitive LFAs and / or any other suitable rapid diagnostic test device.

[0040] A schematic example of a sandwich LFA 170A is shown in FIG. 2A in context. The sandwich LFA 170A (referred to herein as the "LFA") includes a substrate 171 with a sample element 172, a conjugate element 173, a capture element 174, a control element 175, and a wick 176 disposed thereon. The substrate 171 can be of any suitable shape, size, and / or configuration. For example, the substrate 171 can be a rectangular backing card or strip of a certain width and a predetermined length that can provide a sufficient surface area to accommodate the various components of the LFA 170A. The substrate 171 can be made from a semi-rigid polymer designed to provide uniformity and lay-flat characteristics. The substrate 171 can include one or more pressure-sensitive adhesives configured to facilitate the attachment of the various components of the LFA 170A as further described herein.

[0041] As shown, the sample element 172 is generally disposed at one end of the substrate and is configured to receive a sample volume. The sample element 172 provides a surface for receiving a sample of blood and / or other biological fluids for analysis and can be a pad that facilitates the smooth, continuous, and uniform transport of the sample to other components of the lateral flow test strip. The sample element 172 can be of any suitable shape, size, and / or configuration. In some embodiments, the sample element 172 can be configured to prepare the received sample volume for conjugation. For example, the sample element 172 can be configured to perform monocyte depletion, particle (e.g., cell, biomolecule, etc.) immobilization, cell lysis, cell permeabilization, etc. Cell lysis can enable the release of intracellular or membrane-bound proteins such as surface antigen classification 64, which includes intracellular domains, transmembrane domains, and extracellular domains. Further, the sample element 172 can be configured to cleave protein domains from membrane-bound proteins such as extracellular domains from surface antigen classification 64. The conjugate element 173 is disposed adjacent to the sample element 172 in the downstream direction. The conjugate element 173 includes a dry matrix (e.g., a salt-sugar matrix) configured to contain desired bioactive particles. In some embodiments, as described with reference to FIG. 2B, the desired bioactive particles can correspond to two or more target antigens or analytes (i.e., multiplexing). The bioactive particles contained in the matrix include specific antibodies and / or affinity reagents (e.g., DNA aptamers, protein binders, etc.) immobilized on or in the conjugate element 173. The antibodies and / or affinity reagents can be selected based on the target molecule (e.g., antigen or analyte) that the LFA 170A is configured to detect. Further, the antibodies and / or affinity reagents are conjugated directly or indirectly to a molecule configured to enable detection. For example, the antibody can be labeled with a colored particle (e.g., latex having blue color, colloidal gold having red color, and / or any other suitable particle), a fluorescent particle, a magnetic particle, an enzyme for subsequent signal generation, etc.Accordingly, the labeled antibody can bind to the desired antigen or analyte, thereby producing a label or target analyte 177 that can be detected in other portions of the LFA 170A or by other elements.

[0042] The capture element 174 is disposed adjacent to and / or downstream of the conjugate element 173 and includes particles or molecules immobilized within or on the capture element 174. In some embodiments, there can be two or more capture elements 174, as described with reference to FIG. 2B. The particles or molecules can be configured to bind to the labeled analyte 177, thereby capturing or immobilizing the labeled analyte 177 within or on the capture element 174. As the concentration of the captured and / or immobilized labeled analyte 177 increases (e.g., the number of molecules within the capture element 174 increases), the optical density of the detection molecule (e.g., a colored label) also increases. In this way, the LFA 170A is configured to present discrete colorimetric signal lines, regions, or strips indicative of the presence of the target analyte in the sample volume (e.g., a positive test result). In some embodiments, the LFA 170A is configured to provide one or more of a time-resolved fluorescence, electrochemical signal, foaming signal, fluorescence signal, and / or colorimetric signal indicative of the presence of the target analyte in the sample volume.

[0043] The control element 175 is disposed adjacent to and / or downstream of the capture element 174. The control element 175 contains particles or molecules fixed within or on the control element 175. In contrast to the capture element 174, the particles or molecules contained within the control element 175 can be configured to bind to a plurality of different particles, such as, for example, labeled analytes 177, labeled bioactive particles not bound to an antigen. Thus, the control element 175 can bind to labeled particles that are not otherwise immobilized within or on the capture element 174 and / or can be configured to immobilize them otherwise. Thus, the control element 175 can present a colored portion or strip that can be used to indicate that a reaction has occurred and / or that a test has been performed. For example, if the target analyte is not present within the sample volume, it may be desirable to confirm that the assay has been properly performed and that the negative result (absence of a colored strip presented on or by the capture element 174) indicates the status of the sample volume and does not indicate a malfunction of the LFA 170A. The wick 176 is disposed adjacent to and / or downstream of the control element 175 and is configured to absorb or wick portions of the sample that are not immobilized within or on the capture element 174 and / or the control element 175.

[0044] A schematic example of the sandwich LFA 170B for multiplexing is shown in FIG. 2B. The sandwich LFA 170B includes a substrate 171 on which a sample element 174, a conjugate element 173, a first capture element 172', a second capture element 174'', a control element 175, and a wick 176 are disposed. The substrate 171 can be of any suitable shape, size, and / or configuration. For example, the substrate 171 can be a rectangular backing card or strip of a certain width and a predetermined length that can provide a sufficient surface area to accommodate the various components of the LFA 170B. The substrate 171 can be made from a semi-rigid polymer designed to provide uniformity and lay-flat characteristics. The substrate 171 can include one or more pressure-sensitive adhesives configured to facilitate the attachment of the various components of the LFA 170B, as further described herein.

[0045] As shown in the illustration, the sample element 172 is generally disposed at one end of the substrate and configured to receive a sample volume. The sample element 172 provides a surface for receiving a sample of blood and / or other biological fluids for analysis and can be a pad that facilitates the smooth, continuous, and uniform transport of the sample to other components of the lateral flow test strip. The sample element 172 can be of any suitable shape, size, and / or configuration. In some embodiments, the sample element 172 can be configured to prepare the received sample volume for conjugation. For example, the sample element 172 can be configured to perform monocyte depletion, particle (e.g., cell, biomolecule, etc.) immobilization, cell lysis, cell permeabilization, etc. Cell lysis can enable the release of intracellular or membrane-bound proteins such as surface antigen classification 64 that includes intracellular domains, transmembrane domains, and extracellular domains. Further, the sample element 172 can be configured to cleave protein domains from membrane-bound proteins such as extracellular domains from surface antigen classification 64. The conjugate element 173 is disposed adjacent to the sample element 172 in the downstream direction. The conjugate element 173 includes a dry matrix (e.g., a salt-sugar matrix) configured to contain desired bioactive particles. As shown in FIG. 2B, the conjugate element 173 can be configured to provide bioactive particles corresponding to two or more target antigens or analytes (177’, 177’’). The bioactive particles contained in the matrix include specific antibodies and / or affinity reagents (e.g., DNA aptamers, protein binders, etc.) immobilized on or in the conjugate element 173. The antibodies and / or affinity reagents can be selected based on the target molecule (e.g., antigen or analyte) that the LFA 170B is configured to detect. Further, the antibodies and / or affinity reagents are conjugated directly or indirectly to a molecule configured to enable detection. For example, the antibody can be labeled with a colored particle (e.g., latex having blue color, colloidal gold having red color, and / or any other suitable particle), a fluorescent particle, a magnetic particle, an enzyme for subsequent signal generation, etc.Accordingly, the labeled antibody can bind to the desired antigen or analyte, thereby producing a label or target analyte 177 that can be detected in other parts of the LFA170A or by other elements.

[0046] The first capture element 174' and the second capture element 174'' are disposed adjacent to and / or downstream of the conjugate element 173 and include particles or molecules immobilized within or on the first capture element 174' and the second capture element 174''. In some embodiments, the particles or molecules bind to a respective different one of the labeled analytes 177', 177'' such that the labeled analytes 177', 177'' are captured or immobilized within or on a respective one of the first capture element 174' and the second capture element 174''. As the concentration of the captured and / or immobilized labeled analytes 177', 177'' increases (e.g., the number of molecules within the first capture element 174' and the second capture element 174'' increases), the optical density of the detection molecule (e.g., a colored label) also increases. In this way, the LFA170B is configured to present distinct colorimetric signal lines, regions, or strips to indicate the presence of the target analyte in the sample volume. In some embodiments, the LFA170B is configured to provide one or more of time-resolved fluorescence, electrochemical signals, foam signals, fluorescence signals, and / or colorimetric signals to indicate the presence of the target analyte in the sample volume. In some embodiments, the capture elements 174', 174'' are configured to generate one or more of a quantitative signal and a qualitative signal and / or transmit the results of the lateral flow assay to an electronic device (e.g., an electronic device as described in FIG. 21).

[0047] The control element 175 is disposed adjacent to and / or downstream of the capture elements 174', 174''. The control element 175 contains particles or molecules fixed within or on the control element 175. In contrast to the capture elements 174', 174'', the particles or molecules contained within the control element 175 can be configured to bind to a plurality of different particles, such as, for example, labeled analyte 177, labeled bioactive particles not bound to an antigen. Thus, the control element 175 can be configured to bind to labeled particles that are not otherwise immobilized within or on the capture elements 174', 174'', and / or to immobilize them in other cases. Thus, the control element 175 can present a colored portion or strip, or a result configured to be transmitted to an electronic device in another way, which can be used to indicate that a reaction has occurred and / or that a test has been performed. For example, if the target analyte is not present in the sample volume, it may be desirable to confirm that the assay was performed properly and that the negative result (no positive result presented on or by the capture elements 174', 174'') indicates the status of the sample volume and does not indicate a malfunction of the LFA 170B. The wick 176 is disposed adjacent to and / or downstream of the control element 175 and is configured to absorb or wick portions of the sample that are not immobilized within or on the capture elements 174', 174'' and / or the control element 175.

[0048] Assay LFA170A (and / or LFA170B of FIG. 2B) can be used to examine the presence of any suitable target analyte, biomarker, molecule, particle, etc. in a sample volume (e.g., a blood sample or any other suitable sample of body fluid). For example, any of the embodiments described herein can include and / or implement an LFA (e.g., LFA170A, 170B) and / or any other suitable flow-based rapid diagnostic system configured to examine the presence of a specific analyte or biomarker that can provide information used to diagnose a patient condition such as sepsis.

[0049] For example, blood lactate can be a biomarker used in the clinical diagnosis and management of sepsis. In some examples, other biomarker hosts can be used as an alternative to lactate or in addition to lactate to guide clinical decision-making. A non-exhaustive list of suitable biomarkers can include CD64, a type of transmembrane glycoprotein found on the surface of monocytes, macrophages, dendritic cells, and neutrophils; pro-inflammatory cytokines and / or chemokines associated with the hyper-inflammatory phase of sepsis; C-reactive protein and / or procalcitonin (PCT) synthesized in response to infection and inflammation; biomarkers associated with the neutrophil-to-lymphocyte ratio in peripheral blood as a reflection of the balance between systemic inflammation and immunity; biomarkers associated with the activation of neutrophils and / or monocytes (e.g., CD64 as described above); anti-inflammatory cytokines associated with the immunosuppressive phase of sepsis; and / or changes in cell surface markers of monocytes and / or lymphocytes. Pro-inflammatory cytokines and anti-inflammatory cytokines can each include interleukin 6 (IL-6) and interleukin 10 (IL-10) in different examples. In some examples, combinations of pro-inflammatory and anti-inflammatory biomarkers in multiplexed LFAs can be used to identify, for example, patients developing severe sepsis prior to substantial organ dysfunction. In some examples, one or more aptamers can be synthesized to target any other suitable biomarker such as a specific pro-inflammatory biomarker, anti-inflammatory biomarker, and / or any of the biomarkers described herein.

[0050] In some embodiments, LFA170A (and / or LFA170B of FIG. 2B) may be configured to examine the presence of at least a portion of one or more of the biomarkers described above. For example, if the biomarker is a membrane-bound protein comprising at least an intracellular domain, a transmembrane domain, and an extracellular domain, the LFA may be configured to separately detect the intracellular domain, transmembrane domain, and / or extracellular domain of the biomarker, and / or the LFA may be configured to detect the biomarker as a complete (e.g., intact) biomarker.

[0051] Lactate In some implementations, any of the embodiments described herein can be used to detect a lactate biomarker, a PCT biomarker, a CD64 biomarker, and / or any other suitable biomarker described herein that is associated with and / or used to identify sepsis. For example, in some implementations, the rapid test device 170 can be configured to examine the blood lactate concentration in a sample of a body fluid (e.g., blood) using, for example, a portable blood gas analyzer. In other implementations, the rapid test device 170 can be an LFA (e.g., LFA170A, 170B) that is configured to examine a blood (e.g., whole blood, serum, etc.) lactate biomarker (e.g., antigen). For example, the effectiveness of using serum lactate concentration in the diagnosis of sepsis is shown in Table 1 below, which presents the results of a study of acute hospital mortality according to serum lactate concentration in sepsis patients who require vasopressors (e.g., drugs that cause vasoconstriction).

[0052] [Table 1]

[0053] Lactic acid is the end product of anaerobic decomposition of glucose in tissues and can dissociate into lactate, a hydroxy monocarboxylic acid anion that is the conjugate base of lactic acid resulting from the deprotonation of the carboxy group. The generation of lactate in the body occurs when the energy demand by tissues is not met by sufficient aerobic respiration. Lactate can be transported in the blood to the liver, where it is converted to glucose via the Cori cycle. However, without proper clearance of lactate by the liver and kidneys, the accumulated concentration of lactate can lead to lactic acidosis. Clinically, the causes of acidosis can be classified into type A disorders, where tissue oxygenation is reduced, such as in sepsis, and type B disorders, which are caused by certain drugs and / or toxins, especially in association with systemic diseases. Medical evidence suggests that the morbidity and mortality of patients with continuously elevated lactate concentrations are increasing. Excessive lactate in the body can also cause, among other things, bleeding, respiratory failure, trauma, seizures, ischemia, kidney problems, liver disease, tissue hypoxia, shock, blood loss, and anemia. Therefore, lactate monitoring is of utmost importance for diagnosing and evaluating health problems that occur in oxygen-deprived situations (i.e., situations where the lactic acid concentration in the body increases beyond the tolerated value). The lactate concentration in the blood of healthy and unstressed individuals has been reported to be in the range of 0.1 - 1.0 millimolar (mM). In contrast, critically ill individuals, such as those showing severe sepsis or septic shock, can exhibit concentrations higher than 4 mM.

[0054] Lactate can exist as one of two optical isomers, L-lactate and its mirror image, D-lactate. Analytical methods used to detect and quantify lactate include High Performance Liquid Chromatography (HPLC), fluorescence quantification, colorimetric tests, chemiluminescence, and magnetic resonance spectroscopy. These methods can provide accurate results, but they have drawbacks such as time-consuming sample preparation, use of expensive equipment, and the need for trained personnel. As a result, the use of these analytical methods for detecting and quantifying lactate in biological fluids is more suitable for centralized laboratories, and their implementation as point-of-care diagnostic tools may be limited.

[0055] Alternatively, detection of lactate concentration in biological fluids containing blood and / or plasma can be achieved using enzymes. These enzymes can be immobilized on a solid surface or support (e.g., a biosensor) to provide reaction sites that catalyze the lactate chemical reaction by stabilizing the transition reaction state or reducing the activation energy of a specific lactate chemical reaction, generating one or more species that can be monitored to correlate the occurrence of the species with the concentration of lactate. For example, L-lactate can be detected using enzymes such as L-lactate oxidase (LOD) and L-lactate dehydrogenase (LDH). LOD is a globular flavoprotein that can be obtained from various bacterial sources such as Pediococcus, Aerococcus, viridans, and Mycobacterium. The source of LOD can affect the pH range in which the enzyme can exhibit sufficient catalytic activity, which typically shows a pH range of 4 - 9. LOD, which is a member of the flavin mononucleotide (FMN) family, uses FMN as a cofactor to catalyze the oxidation of hydroxy acids in the reaction of LOD with glycolic acid oxidase, L-lactate, monooxygenase, flavocytochrome b2, long-chain α-hydroxylate oxidase, and L-mandelate dehydrogenase. To detect the presence of L-lactate, LOD can be immobilized on a solid support and exposed to biological fluids such as blood and plasma. LOD can catalyze the oxidation of L-lactate to pyruvic acid (PA) in the presence of dissolved oxygen, generating reduced LOD and hydrogen peroxide (H2O2) as by-products. The hydrogen peroxide generated from lactate oxidation can be accurately quantified by secondary chemical reactions and / or electrochemical reactions. For example, the hydrogen peroxide generated during the oxidation of lactate in the presence of the LOD enzyme can be electrochemically reduced or oxidized to generate an electrical signal that can be monitored by an electrode. The reduced LOD enzyme can then be re-oxidized in a second reaction step on the electrode as shown in the following reaction scheme.

[0056] [Chemistry]

[0057] Similar to LOD, the LDH enzyme can be used to detect and quantify the presence of L-lactic acid in various biological fluids. LDH is a tetrameric protein that can be found in animals, plants, and prokaryotes. LDH is latent throughout tissues and is released during tissue damage. The LDH enzyme contains five different isoenzyme forms that are distinguished by slight structural differences. Depending on the source, the LHD enzyme is known to be stable over a relatively narrow pH range of 5 - 8, more specifically, a pH range of approximately 7.2 - 7.4. LDH can also catalyze the reaction from L-lactic acid to pyruvic acid (PA) via its cofactor, nicotinamide adenine dinucleotide (NAD), which can exist in an oxidized form (NAD + ) and a reduced form (NADH). During the reaction, LHD converts L-lactic acid to pyruvate (PA) and converts NAD + to NADH. Subsequently, the detection of lactate using the LDH enzyme can be achieved by a secondary reaction as described above for the detection of L-lactic acid using the LOD enzyme. For example, NADH can be electrochemically oxidized under the influence of an applied potential generated using an electrode as shown in the following reaction scheme, and the resulting current is proportional to the L-lactic acid concentration.

[0058] [Chemistry]

[0059] Detecting lactate in a biological fluid via lactate oxidase using an enzyme, as described above, relies on the conversion of lactate to one or more by-products such as NADH and hydrogen peroxide (H2O2) that can be accurately quantified by a secondary reaction. The secondary reaction often involves an electrochemical conversion that occurs at the surface of the electrode and generates a transient current proportional to the amount of lactate present in the sample (e.g., electrochemical techniques for lactate sensing). Alternatively, the by-products of the enzymatic reaction of lactate can be quantified by light transfer processes (e.g., electrochemiluminescence and fluorescence techniques for lactate sensing), as further described herein.

[0060] Biosensors that rely on electrochemical techniques for the detection of lactate (i.e., electrochemical biosensors) use an enzyme immobilized on a support substrate located near or in the vicinity of the electrode surface. The performance characteristics of an electrochemical biosensor can vary widely depending on the enzyme source, environmental conditions including pH and temperature, the method used to immobilize the enzyme on the biosensor, the chemical nature of the matrix or support used to immobilize the enzyme, and / or the electron transfer mechanism. The enzyme can be immobilized according to different methods, and the reactivity of the enzyme depends on the interaction between the enzyme and the support, the nature of the enzyme, and the presence of adsorbed species, mediators, and additives. Common enzyme immobilization techniques include physical adsorption, entrapment behind a dialysis membrane or polymer film, covalent bonding via a cross-linking agent, and incorporation into the bulk of a carbon composite matrix.

[0061] Problems associated with enzyme immobilization include reproducibility, stability, and inactivation due to the generation and / or accumulation of inhibitors and / or contaminating species. For example, LOD enzymes immobilized via physical adsorption on a biosensor containing an Au electrode may show a 50% loss of stability after only 1 month of storage, while LOD enzymes immobilized in mesoporous silica using a polymer matrix of polyvinyl alcohol (PVA) may show 98% of their initial activity after 9 months. As a result, to develop a sensor for detecting lactate using LOD enzymes, it is necessary to identify appropriate immobilization techniques, suitable matrix supports, and use and / or storage environmental conditions such that the enzyme activity or shell life can be maintained over a long period.

[0062] Electrochemical biosensors for lactate detection typically include devices with two or three electrode sensing platforms. Accurately measuring lactate often involves the use of a reference electrode (commonly made of Ag / AgCl2) held in close proximity to the working electrode to maintain a stable known potential. The working electrode functions as a transducer, and the counter electrode establishes a path for current to flow in response to potential changes at the working electrode. Common techniques for measuring the electrical signals generated during lactate detection include cyclic voltammetry, amperometry, and potentiometry. Electrochemical biosensors can provide high sensitivity, a wide linear range, and a rapid response. However, the use of electrochemical biosensors presents limitations due to complex experimental setups, passivation of the system by fouling agents, and signal reduction and interference due to competing reactions. For example, the electrochemical quantification of hydrogen peroxide (H2O2) generated during the enzymatic oxidation of L-lactate on LOD enzymes requires a high oxidation potential, thereby causing interference by other electrochemically oxidizable species.

[0063] A lateral flow assay (LFA) configured to assay a blood (e.g., whole blood, serum, etc.) lactate biomarker (e.g., antigen) provides an alternative tool for facilitating and / or assisting in the diagnosis of sepsis. As described above with reference to FIG. 2A, the LFA can be performed on a strip that includes one or more components assembled on a plastic backing laminate or substrate 171. The components of the LFA configured to quantify lactate in blood and / or other biological fluids can at least include a sample element 172 and a conjugate element 173.

[0064] The sample element 172 can be a pad that provides a surface for receiving a sample of blood and / or other biological fluids for analysis, and as described above, facilitates the smooth, continuous, and uniform transport of the sample to the other components of the lateral flow test strip. The sample element 172 can be of any suitable shape and / or size. In some embodiments, the shape of the sample element 172 can be a rectangular strip configured to adsorb and receive a sample of a volume of blood and / or other biological fluids. In other embodiments, the shape of the sample element 172 can be a rectangular strip, and one of the ends of the strip can include a region having a dimension larger than the width of the strip to facilitate pipetting a sample of a volume of blood and / or other biological fluids. For example, the sample element 172 can be a rectangular strip that includes a circular region attached to one of the ends of the strip. The circular region of the sample element 172 can provide a larger surface area for receiving a sample of blood and / or other biological fluids via a micropipette. Alternatively, in some embodiments, the sample element 172 can include a large-diameter circular-shaped region with various rectangular strips originating radially from the center of the circular-shaped region. Each rectangular strip can facilitate the transport of a portion of the sample of blood and / or other biological fluids to the other components of the lateral flow test strip for simultaneous detection (i.e., multiplexing) of multiple biomarkers and / or for replicating the assay for verification purposes.

[0065] The sample element 172 can be disposed on the surface of a plastic backing laminate to provide mechanical support to the LFA. In some embodiments, the sample element 172 can include an adhesive coated on one surface of the sample pad to facilitate attachment to the plastic backing laminate. The shape and dimensions of the sample element 172 can be predetermined such that the sample element can be disposed on the plastic backing laminate. The thickness of the sample element 172 can be selected to facilitate adhesion of the sample element 172 to the plastic backing laminate while maintaining the mechanical structure of the pad. Further, the thickness of the sample element 172 can be selected to accommodate a large amount of blood and / or other biological fluids and prevent oversaturation of the sample on the pad and cross-flow to the plastic backing laminate. For example, in some embodiments, the thickness of the sample element 172 can be from 0.18 mm to 0.34 mm.

[0066] The sample element 172 can be made from cellulose, nitrocellulose, glass fiber, and / or any other suitable material. In some embodiments, the sample element 172 can be made from a cellulose membrane and / or chromatography paper configured to promote a linear flow rate of about 3-5 mm / min. The sample element 172 can also include one or more chemical reagents configured to pretreat the sample prior to transport of the sample to other downstream components. In some embodiments, the surface of the sample element 172 can be impregnated with an aqueous buffer that provides an environment having a controlled pH. In some embodiments, the surface of the sample element 172 can be impregnated with a buffer solution including, but not limited to, phosphate-buffered saline (PBS), 2-(N-morpholino)ethanesulfonic acid (MES), tris(hydroxymethyl)aminomethane (TRIS), piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES), 3-(N-morpholino)propanesulfonic acid (MOPS), 2-[4-(2-hydroxyethyl)piperazine-1-yl]ethanesulfonic acid (HEPES), [tris(hydroxymethyl)methylamino]propanesulfonic acid (TAPS), N-cyclohexyl-2-aminoethanesulfonic acid (CHES), and / or N-cyclohexyl-3-aminopropanesulfonic acid (CAPS).

[0067] In some embodiments, the sample element 172 can include one or more components configured to capture and separate species present in blood and / or other biological fluids that can cause interference with the LFA assay. For example, in some embodiments, the sample element 172 can include one or more regions configured to separate red blood cells present in a blood and / or other biological fluid sample. In some examples, the region configured to separate blood cells can be one or more discrete pads that can be disposed on the sample element 172. In other embodiments, the blood separation region can be a pad located adjacent to the sample element 172. In some examples, the blood separation pad can include one or more layers such as a polyester matrix and a composite matrix, and these layers are designed to have an asymmetric morphology with different porosities and pore size distributions that facilitate the capture of the cellular components of blood (i.e., red blood cells, white blood cells, and platelets) in larger pores while allowing plasma to flow downstream through smaller sized pores.

[0068] The conjugate element 173 of the LFA for the detection and quantification of lactate from a blood and / or other biological fluid sample can be a pad located adjacent and downstream of the sample element 172. The conjugate element 173 can include a dry matrix (e.g., a salt-sugar matrix) that contains bioactive species capable of reacting with lactate and generates species that can be detected by colorimetry, as further described herein. The conjugate element 173 can be configured to accommodate one or more bioactive species that can be released upon contact with the mobile liquid sample deposited on the upstream sample element 172. As described above with reference to the sample element 172, the conjugate element 173 can be a pad of any suitable shape and / or size. In some embodiments, the shape of the conjugate element 173 can be a strip having a size and / or shape that is substantially similar to the size and / or shape of the sample element 172. In some embodiments, the conjugate element 173 and the sample element 172 can be made from a single pad, disposed at opposite ends thereof, and optionally attached to the surface of a plastic backing laminate to provide mechanical support to the LFA. In yet another embodiment, the conjugate element 173 and the sample element 171 can be made from a single pad that includes a rectangular strip, wherein the first end of the strip includes a region having a dimension larger than the width of the strip to provide a region for accommodating bioactive species for lactate oxidation and colorimetric detection, and the second end of the strip on the opposite side of the first end includes a region having a dimension larger than the width of the strip to provide a region for accommodating a volume of a sample of blood and / or other biological fluid. Alternatively, in some embodiments, the conjugate element 173 can include a plurality of rectangular strips (FIG. 2B) connected to a radially larger circular-shaped region, the larger circular-shaped region being configured to accommodate the sample element 171. In this configuration, each conjugate element 173 can facilitate the detection (i.e., multiplexing) of a plurality of biomarkers present in a portion of a sample of blood and / or other biological fluid, and / or the replication of assays for validation purposes.

[0069] The conjugate element 173 can include a dry matrix configured to include a desired bioactive species for the detection and quantification of lactate in a sample of blood and / or other biological fluids. For example, in this embodiment, the matrix of the conjugate element 173 can include both a detection enzyme and a quantification enzyme. The detection enzyme exhibits high activity and selectivity towards the catalytic oxidation of lactate and is configured to generate one or more by-products that can be monitored by a secondary chemical reaction to quantify the concentration of lactate present in the sample. For example, in some embodiments, the matrix of the conjugate element 173 can include a detection enzyme such as L-lactate oxidase (LOD). In other embodiments, the matrix of the conjugate element 173 can include other suitable detection enzymes such as L-lactate dehydrogenase (LDH). The one or more detection enzymes can be loosely deposited on the surface of the conjugate element 173 pad such that they can be dissolved in a certain volume of the sample of blood and / or other biological fluids flowing from the sample element 172.

[0070] The quantification enzyme exhibits high activity and selectivity towards the stoichiometric conversion of one or more species generated during the enzymatic oxidation of lactate and is configured to generate a signal that can be quantified. In some embodiments, the dry matrix can include one or more heme-containing enzymes such as catalase and / or peroxidase that can catalyze the redox reaction with a hydroperoxide such as hydrogen peroxide (H2O2) generated during lactate oxidation. The heme-containing enzyme can be, for example, horseradish peroxidase, which can catalyze the redox reaction of hydrogen peroxide (H2O2) and 3,3'-diaminobenzidine (3,3'-diaminobenzidine, DAB) and generate a dark brown insoluble product that can be detected and quantified by colorimetric analysis.

[0071] Although LFA170A and LFA170B were described above as also including at least one capture element 174, a control element 175, and a wick 176, in this embodiment, the detection of lactate can be performed, for example, on or at the conjugate element 173. Thus, the LFA need not include separate capture, control, and / or wick elements.

[0072] In some embodiments, for example, the LFA is coupled to an optical device such as a CMOS or CCD camera configured to collect an image of a 3,3'-diaminobenzidine (DAB) dark brown precipitate resulting from oxidation by hydrogen peroxide to determine the concentration of lactate originally present in the sample. For example, in some embodiments, the conjugate element 173 of the LFA can be imaged by a camera of a peripheral device such as a smartphone or a dedicated optical detector, and the intensity of the image can be analyzed by image software to estimate the concentration of the DAB precipitate, the concentration of hydrogen peroxide, and thus the concentration of lactate originally present in the sample. In some embodiments, the concentration of lactate present in the sample can be determined by (1) recording an image of the DAB brown precipitate, (2) calculating the grayscale mode value using image processing software, and (3) correlating the concentration of lactate present in a sample of known lactate content with the grayscale mode value. The range of grayscale mode values that an image can assume is 0 to 255, with values closer to zero corresponding to darker images and values closer to 255 corresponding to brighter images.

[0073] A lateral flow assay (LFA) configured to detect lactate in blood and / or other biological fluids can address certain drawbacks observed in lactate detection approaches that rely on electrochemical reactions to quantify the amount of hydrogen peroxide (H2O2) generated during lactate oxidation. As described above, the enzymatic reaction between hydrogen peroxide (H2O2) and 3,3'-diaminobenzidine (DAB) produces a brown precipitate that is insoluble in blood and / or biological fluid samples and can be quantified by optical methods such as colorimetry. Furthermore, the reaction between hydrogen peroxide and DAB proceeds under pH and temperature conditions similar to those required for lactate oxidation. Thus, the use of additives in the dry matrix of the LFA can protect both the detection and quantification enzymes from degradation, facilitating storage for as long as nine months, as further described herein. In contrast, electrochemical methods for detecting and quantifying lactate typically require the use of high oxidation potentials to convert hydrogen peroxide into an electrical signal. Such potentials can frequently cause interfering reactions with other electro-oxidizable species present in blood and / or biological fluid samples, resulting in inaccurate results. Additionally, the immobilization of enzymes on solid surfaces can present several challenges, including the need for complex and / or time-consuming fabrication and characterization methods and a decrease in enzyme stability during storage.

[0074] In some embodiments, the detection enzyme and the quantification enzyme can be included in a dry matrix in the presence of one or more chemical reagents and / or stabilizing additives configured to preserve the activity and stability of the enzyme during storage and during the oxidation of lactate in blood and / or other biological fluid samples. For example, the dry matrix can contain a weak acid or base (e.g., a buffer) that can be dissolved in blood and / or other biological fluid samples, dissociate in the sample, establish an equilibrium between their acidic species and their conjugates, and maintain the pH of the sample within a range of values at which the enzyme exhibits high catalytic activity. Some embodiments can include one or more buffers such as phosphate buffered saline (PBS), 2-(N-morpholino)ethanesulfonic acid (MES), tris(hydroxymethyl)aminomethane (TRIS), piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES), 3-(N-morpholino)propanesulfonic acid (MOPS), 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES), [tris(hydroxymethyl)methylamino]propanesulfonic acid (TAPS), N-cyclohexyl-2-aminoethanesulfonic acid (CHES), and / or N-cyclohexyl-3-aminopropanesulfonic acid (CAPS) in the dry matrix.

[0075] In some embodiments, the dry matrix can include a polysaccharide such as chitosan, an antibacterial activity, and a non-toxic biocompatible biopolymer that can provide antioxidant activity for long-term preservation of the chemical integrity of the enzyme. In some embodiments, the chitosan stabilizer can be accompanied by one or more reagents configured to increase the solubility of chitosan in a sample of blood and / or other biological fluids. For example, in some embodiments, the dry matrix can include chitosan and a weak organic acid such as formic acid, acetic acid, and / or propionic acid suitable for increasing the solubility of chitosan in a certain volume of blood and / or biological fluid. In some embodiments, the dry matrix can include a combination of additives including chitosan, acetic acid, and / or a buffer, and the combination of additives is adsorbed on the surface of conjugate 173 and configured to be dissolved in a certain volume of blood and / or biological fluid transported from sample element 170.

[0076] As described above, a lateral flow assay (LFA) configured to assay lactic acid in blood and / or other biological fluids can detect lactic acid present in various samples including buffer solution, serum, plasma, and / or whole blood. More specifically, in some embodiments, the LFA can exhibit a dynamic range of detectable lactate from 2 to 6 (mM), as well as a sensitivity of 0.5 mM lactate or more in buffer and / or serum samples. In some embodiments, the LFA can exhibit cut-off lactate concentrations of 2 mM and 4 mM in buffer / serum. The total time required to obtain a lactate result using an LFA configured for lactate detection can be about 10 minutes. An LFA configured for lactate detection, when subjected to an accelerated degradation study at 37 °C, can mainly degrade during the first week of the assay and remain relatively stable over time. More specifically, an LFA configured for lactate detection can remain stable up to 4 weeks at 37 °C, showing a small change in signal response, supporting the idea that the LFA assay can remain viable over a long period.

[0077] Procalcitonin In some implementations, the rapid test device 170 can be an LFA (e.g., LFA 170A, 170B) configured to test for PCT biomarkers. For example, Table 2 below shows the effectiveness of using serum PCT biomarker concentrations in blood in the diagnosis of sepsis. Table 2 shows the results of a study on the diagnosis of sepsis, severe sepsis, and septic shock by serum PCT measurement.

[0078]

Table 2

[0079] Procalcitonin (PCT) is an 116 - amino acid peptide with an approximate molecular weight MW of 14.5 kDa and belongs to the calcitonin family of peptides. The PCT molecule consists of three parts: an amino - terminal (57 amino acids), pro - calcitonin (33 amino acids), and the calcitonin carboxyl - terminal peptide 1 (CCP - 1), also known as katacalcin (21 amino acids). PCT is a precursor hormone of calcitonin and cannot be detected in healthy individuals because the peptide is not released into the blood in the absence of systemic inflammation. However, in the case of sepsis caused by bacterial infection, PCT synthesis is induced in tissues and thus becomes detectable in the blood. The production of PCT can be caused by bacterial toxins such as endotoxin and cytokines (e.g., interleukin (IL) - 1β, interleukin - 6, and tumor necrosis factor (TNF) - α). PCT levels can increase rapidly between 2 hours and 6 hours after bacterial infection and reach a peak within 6 hours to 24 hours. In addition to bacterial infection, some fungal and parasitic infections are associated with the release of PCT into the bloodstream. Further conditions that result in high levels of PCT in the body include recent major surgery, severe trauma, severe burns, prolonged cardiogenic shock, and chronic kidney disease.

[0080] In some extrathyroidal tissues, when there is no ability to cleave PCT into its mature form, calcitonin, PCT can accumulate in the blood. As a result, PCT can be used in patients suffering from sepsis. It can be used as a biomarker with a relatively high discrimination between bacterial and viral inflammation. Furthermore, PCT levels can be related to and / or indicative of the severity of a bacterial infection. In the case of sepsis, a rapid diagnosis of a bacterial infection reduces the risk of antibiotic resistance or unnecessary or inappropriate use of antibiotics that can increase toxic side effects in the patient.

[0081] Conventional approaches for diagnosing sepsis caused by bloodstream infections involve culturing blood, urine, and cerebrospinal fluid from bronchial fluid specimens. These testing approaches typically can take 24 to 48 hours to obtain results and, in many cases, can facilitate the identification of pathogens and provide information regarding the type of microorganism and its susceptibility to antibiotics. However, when no positive cultures are present, clinical symptoms may appear and treatment based on false-negative results may be administered. Coupled with the half-life of PCT (25 to 30 hours) and the specificity of PCT for bacterial infections and the virtual absence of PCT in healthy individuals, PCT is a suitable biomarker for bacterial infections.

[0082] PCT can be quantified by immunoassays based on the sandwich ELISA principle. In such immunoassays, an antibody-procalcitonin-antibody complex is formed and quantified by one or more measurement techniques including chemiluminescence, enzyme, fluorescence, and turbidimetric immunoassays. For example, a chemiluminescent assay for PCT uses a two-step sandwich approach. In this method, an anti-PCT monoclonal antibody conjugated with alkaline phosphatase is added to the patient sample in the presence of a reagent buffer. After incubation, paramagnetic particles coated with the monoclonal anti-PCT antibody are added to the assay. PCT binds to the paramagnetic particles, while the anti-PCT antibody in solution reacts with different antigenic sites of the PCT molecule. The particles are separated from the unbound material by a magnet. A chemiluminescent substrate is added to the assay, and the light generated by the reaction is measured using a luminometer, where photon generation is proportional to the PCT concentration in the sample.

[0083] Alternatively, PCT can be measured using a quantitative homogeneous assay (BRAHMS, Hennigsdorf, Germany) based on Time Resolved Amplified Cryptate Emission (TRACE) technology. The assay involves directing a nitrogen laser 337 nm beam at a sample containing PCT and two fluorescently labeled antibodies that recognize different epitopes of the PCT peptide. Exposure to the laser excitation results in non-radiative energy transfer between the donor molecule and the acceptor molecule. The donor molecule emits a long-lived fluorescent signal at 620 nm, and the acceptor molecule emits a short-lived signal at 665 nm. When both the donor molecule and the acceptor molecule are brought into proximity by binding to PCT, the resulting signal is amplified at 665 nm and persists for several microseconds. This is long enough to be detected after the decay of the background fluorescence common in biological samples.

[0084] A lateral flow assay (LFA) PCT biomarker (e.g., an antigen) configured to examine blood (e.g., whole blood, serum, etc.) provides an alternative tool for the diagnosis of sepsis. As described above with reference to FIG. 2A, the LFA can be implemented on a strip having one or more components assembled on a substrate 171. The components of the LFA configured to detect and quantify PCT in blood and / or other biological fluids can include a sample element 172, a conjugate element 173, a capture element 174, a control element 175, and a wick 176.

[0085] The substrate 171 can be a backing laminate or a backing card configured to provide mechanical support to the components of the LFA, as described above. The substrate 171 can be of any suitable shape, size, and / or configuration, as described above. For example, the substrate 171 can be a rectangular backing card or strip of a certain width and a predetermined length that can provide a sufficient surface area to accommodate the various components of the LFA. The substrate 171 can be made from a semi-rigid polymer designed to provide uniformity and lay-flat characteristics. The substrate 171 can include one or more pressure-sensitive adhesives configured to facilitate the attachment of the various components of the LFA, as further described herein.

[0086] The sample element 172 can be a pad that provides a surface for receiving a sample of blood and / or other biological fluids for analysis and facilitates the smooth, continuous, and uniform transport of the sample to other components of the lateral flow test strip. The sample element 172 can be of any suitable shape and / or size. In some embodiments, the shape of the sample element 172 can be a rectangular strip configured to adsorb and receive a volume of sample of blood and / or other biological fluids. The sample element 172 can be disposed on the surface of the substrate 171 to provide mechanical support to the LFA. In some embodiments, the sample element 172 can include an adhesive coated on one surface of the sample pad to facilitate attachment to a plastic backing laminate. The shape and dimensions of the sample element 172 can be predetermined such that the sample element can be disposed on the plastic backing laminate. The thickness of the sample element 172 can be selected to facilitate adhesion of the sample element 172 to the plastic backing laminate while maintaining the mechanical structure of the pad. Further, the thickness of the sample element 172 can be selected to accommodate a large volume of blood and / or other biological fluids and prevent oversaturation of the sample on the pad and lateral flow to the plastic backing laminate. The sample element 172 can be made of cellulose, nitrocellulose, glass fiber, and / or any other suitable material.

[0087] The conjugate element 173 of the LFA for the detection and quantification of PCT from blood and / or other biological fluid samples can be a pad located adjacent and downstream of the sample element 172, as shown in FIG. 2A. The conjugate element 173 can be of any suitable shape, size, and / or configuration. In some embodiments, the shape of the sample element 172 can be a rectangular strip of a width similar to that of the sample member 171, disposed on the surface of the substrate 171 to provide mechanical support to the LFA. The conjugate element 173 can include a dry matrix (e.g., a salt-sugar matrix) containing bioactive particles and additives. The bioactive particles contained in the matrix include specific antibodies and / or affinity reagents (e.g., DNA aptamers, protein binders, etc.) immobilized on or in the conjugate element 173. For example, in some embodiments, the surface of the sample element 172 can be impregnated with an aqueous buffer that provides an environment having a controlled pH. In some embodiments, the surface of the sample element 172 can be impregnated with a buffer solution including, but not limited to, borate buffer solution, phosphate buffered saline (PBS), 2-ethanesulfonic acid (MES), tris(hydroxymethyl)aminomethane (TRIS), piperazine-N,N'-bis(PIPES), 3-morpholinopropane-1-sulfonic acid (MOPS), 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES), [tris(hydroxymethyl)methylamino]propanesulfonic acid (TAPS), N-cyclohexyl-2-aminoethanesulfonic acid (CHES), and / or N-cyclohexyl-3-aminopropanesulfonic acid (CAPS).

[0088] The dry matrix of the sample element 172 can include one or more surfactants used as wetting agents to solubilize polar species present in the sample. For example, in some embodiments, the dry matrix of the conjugate element 173 can include nonionic surfactants such as glycidol, tergitol, ethoxylated and alkoxylated fatty acids, ethoxylated amines, alkyl and nonylphenol ethoxylates, ethoxylated sorbitan esters, castor oil ethoxylates, etc. The dry matrix can include one or more biocides configured to promote the extended shelf life of the LFA by inhibiting a wide range of microorganisms. The biocide can be formulated at a low concentration in the dry matrix of the conjugate element 173 to minimize and / or avoid potential health hazards, toxicological problems, and disposal problems. For example, in some embodiments, the dry matrix can include other commercially available preservative formulations such as 5-chloro-2-methyl-4-isothiazolin-3-one (CMIT), 2-methyl-4-isothiazolin-3-one (MIT), proprietary glycols, modified alkyl carboxylates, and / or proclin 300 (trademark). In some embodiments, the dry matrix can include one or more surfactants, or any amphiphilic molecule (e.g., tween 20, Triton X, octyl thioglucoside, etc.) that can be used for protein solubilization.

[0089] The dry matrix of the conjugate element 173 can include one or more antibodies and / or affinity reagents conjugated to molecules configured to enable detection. In some aspects, the dry matrix can include one or more detection antibodies that can bind to PCT and exhibit high stability. For example, in some aspects, the detection antibody can include procalcitonin human antibodies including monoclonal anti-PCT antibody 14A2cc, monoclonal anti-CT antibody 796, PP3, etc.

[0090] The detection antibody can be immobilized on one or more colored particles (e.g., latex having blue color, colloidal gold having red color, and / or any other suitable particles), fluorescent particles, magnetic particles, or any other suitable particles that can be used for the capture and quantification of PCT in a sample of blood and / or other biological fluids. In some embodiments, the detection antibody can be immobilized on gold nanoparticles. The gold nanoparticles and / or gold nanoshells can be functionalized with an antibody that exhibits specific binding activity to a particular region of the PCT molecule (e.g., bioconjugation). During bioconjugation, the surface of the gold nanoparticles can be functionalized with the detection antibody using physical methods that rely on physical interactions between the detection antibody and the surface of the gold nanoparticles, such as ionic interactions, hydrophobic interactions, and / or donor bonds. The physical interaction is caused by the spontaneous absorption of the antibody onto the surface of the gold nanoparticles. In the case of ionic interactions, the positively charged groups in the detection antibody are attracted to the negatively charged surface of the gold nanoparticles. Hydrophobic interactions occur between the hydrophobic portion of the detection antibody and the metal surface.

[0091] Advantages of functionalizing gold nanoparticles using detection antibodies via physical methods include ease of fabrication, simplicity, low cost, rapid production, and minimal use of additives and / or chemicals that can cause harmful toxic effects. However, certain drawbacks of physical methods can include the use of large amounts of detection antibodies in the preparation of functionalized gold nanoparticles, random orientation of the detection antibodies, and relatively easy replacement of the detection antibodies by other molecules with similar properties. These drawbacks often result in high assay variability and low PCT capture ability due to the low specificity of the binding mode on the gold nanoparticles. For example, the conjugation of an antibody to the surface of a gold nanoparticle can proceed via non-specific binding sites that may block regions of the antibody suitable for PCT capture. For instance, in some examples, the antibody can be physically adsorbed onto the surface of the gold nanoparticle by interactions between the constant domain present in the heavy chain and the surface of the nanoparticle. In this configuration, the antigen-binding site of the antibody may be partially available for interaction with the PCT. In other examples, the antibody can be physically adsorbed onto the surface of the gold nanoparticle by interactions between the antigen-binding site and the PCT antigen-binding site that exclude interaction with the PCT.

[0092] Alternatively, in some embodiments, the antibody can be conjugated to the gold nanoparticles by a chemical method that includes covalent bonding (e.g., chemisorption via thiol derivatives, bifunctional linkers, and / or adapter molecules). Direct functionalization of gold nanoparticles with thiol derivative groups can be achieved by forming strong bonds on the surface of the particles by a chemical reaction between gold atoms and sulfur atoms. For example, a thiol-functionalized antibody can be directly attached to the gold nanoparticles. However, this approach presents challenges such as potentially compromising the stability of the nanoparticles and the use of reaction conditions that may require stringent conditions.

[0093] The embodiments, implementations, and / or methods described herein can overcome these limitations, for example, by including the use of other groups that can be attached to the surface of gold nanoparticles using a bifunctional linker that provides specific functionalization on the surface of the gold nanoparticles. For example, carboxylated polyethylene glycol (PEG) molecules functionalized with thiol groups (PEG-SH) can be used to functionalize the surface of gold nanoparticles. The PEG molecules functionalized on the gold nanoparticles can also include carboxyl end groups. Such carboxyl end groups can be modified in a coupling chemical reaction involving water-soluble carbodiimide (EDC) and N-hydroxy-succinimide (NHS) compounds to generate reactive functional groups that bind to the primary amine groups in antibody molecules. The water-soluble carbodiimide reacts with the carboxylic acid moiety in the PEG containing the gold nanoparticles to generate an intermediate active group, which reacts with the N-hydroxy-succinimide compound to form a reactive ester group. The primary amine groups in the antibody react with the ester group formed on the surface of the gold nanoparticles when in direct contact with the antibody. This reaction is designed to generate an amide bond for attaching the antibody to the gold nanoparticles without adding a spacer molecule between the gold nanoparticles and the antibody.

[0094] The capture element 174 of the LFA for the detection and quantification of PCT from blood and / or other biological fluid samples can be a pad disposed adjacent to and / or downstream of the conjugate element 173, which contains particles or molecules immobilized within or on the capture element 174. As described above with reference to FIG. 2, the particles or molecules can be configured to bind to a detection antibody conjugated to the colored particles described above with reference to the conjugate element 173 as a downstream flow of a volume of blood and / or other biological fluid sample. In some embodiments, the capture element 174 can include a capture antibody immobilized and / or chemically bound to the surface of the capture element 174. The capture antibody can be configured to interact with the detection antibody to capture the PCT bound to the detection antibody, resulting in a localized accumulation of the detection antibody and their conjugated colored particles. In some embodiments, the capture antibody can be adsorbed onto the surface of the capture element 174.

[0095] As described above with respect to the detection antibody, the capture antibody can include a procalcitonin human antibody including monoclonal anti-PCT antibody 14A2cc, monoclonal anti-CT antibody 796, PP3, etc. The immobilized capture antibody can be configured to bind to the PCT molecules pre-bound to the detection antibody (and their conjugated colored particles) at the conjugate element 173. As a result, upon exposing the capture element 174 to a sample of blood and / or other biological fluid containing PCT that has been previously flowed through the conjugate element 173, it is possible to accumulate the associated colored particles that capture the antibodies bound to the PCT molecules present in the sample. Such an accumulation of colored particles on the capture element 174 can be recorded and quantified by one or more optical methods to determine the concentration of PCT in the sample. For example, in some embodiments, the colored particles accumulated on the capture element 174 can be determined by a standard lateral flow reader such as the commercially available Leelu reader (LUMOS diagnostics) configured to detect colored particles that provide sufficient suitable optical sensitivity and dynamic range to cover a wide range of concentrations.

[0096] The LFA can include a control element 175 for capturing detection antibodies that would otherwise not be captured by the capture element 174. In other embodiments, the LFA need not include a control element. The wick 176 of the LFA configured for the detection and quantification of PCT from a blood and / or other biological fluid sample can be a pad disposed adjacent to and / or downstream of the capture element 174 (or the control element 175 if included). As described with reference to FIG. 2A, the wick 176 can be configured to absorb or wick portions of the sample that are not immobilized within and / or on top of the capture element 174 (and / or the control element 175 if included).

[0097] As described above, the LFA configured to assay for PCT in blood and / or other biological fluids can detect PCT present in a variety of samples including buffered solutions, serum, plasma, and / or whole blood. More specifically, in some embodiments, the LFA can exhibit a dynamic range of detectable PCT from 0.2 ng / mL to 2 ng / mL in buffer and serum, as well as a sensitivity of 0.1 ng / mL or greater. In some embodiments, the LFA can exhibit cut-off PCT concentrations of 0.2 ng / mL and 0.5 ng / mL in buffer / serum. The LFA configured for PCT detection, for which the total time required to obtain results using the LFA can be about 10 minutes, can remain relatively stable without significant conjugate release and flow through the lateral flow strip during an accelerated stability test conducted at 37°C.

[0098] Although the testing of serum lactate and / or serum PCT concentrations has been described above, it should be understood that testing of blood (e.g., whole blood or other suitable portions of blood) will yield similar or substantially the same results. Although the lactate biomarker and the PCT biomarker have been described above, it should be understood that those biomarkers are presented by way of example only and are not limiting. In some implementations, rapid test devices (e.g., LFA170A, 170B) can be configured to test for any suitable biomarker associated with and / or indicative of sepsis, such as CD64 (described below), and / or any other infectious or disease condition. Further, it should be understood that rapid test devices 170 and / or LFA170A, 170B (and / or any other suitable flow-based assay) can be used in conjunction with any of the fluid transfer devices described herein with reference to specific embodiments.

[0099] CD64 In some implementations, rapid test device 170 can be an LFA (e.g., LFA170A of FIG. 2A and / or LFA170B of FIG. 2B) configured to test for the CD64 biomarker.

[0100] When activated, neutrophils express Fcγ receptor I, also known as the CD64 antigen. Human CD64 is a transmembrane glycoprotein (72-kDa) that, together with Fcγ receptor II (CD32) and Fcγ receptor III (CD16), constitutes the large immunoglobulin (Ig) superfamily. Human CD64 contains an intracellular domain, a transmembrane domain, and an extracellular domain. Human CD64 binds monomeric IgG (for IgG1, IgG3, and IgG4) with high affinity in the nanomolar range. Compared to other human Fcγ receptors, the affinity of human CD64 for monomeric Ig is 10- to 100-fold higher than that of the low-affinity Fcγ receptor II or Fcγ receptor III family of receptors, which have insufficient interaction with monomeric IgG with binding affinities in the micromolar range.

[0101] CD64 is constitutively expressed on monocytes and macrophages and can also be found on neutrophils together with interferon gamma (IFN-γ) and granulocyte colony stimulating factor (G-CSF). CD64 is also expressed together with Fcγ receptor 2 on the bone marrow-derived cell lines HL-60, THP-1, and U937. Treatment with IFN-γ has been shown to upregulate CD64 expression in U937 cells 4- to 5-fold to levels of 60,000 receptors per cell. Cytokine stimulation induces rapid clustering of CD64 on the cell membrane, promoting the rapid binding and internalization of immune complexes, including de novo protein expression of new CD64 molecules to facilitate complex binding. This contributes to the inflammatory response by essentially causing the release of tumor necrosis factor alpha (TNF-α) and IL-6, superoxide production, antigen presentation to T cells, or lysis of antibody-coated cells.

[0102] In cases of inflammation such as during sepsis, CD64 expression is rapidly upregulated on neutrophils. The intensity of cytokine stimulation during sepsis is directly correlated with the stepwise increase in CD64 expression. Many meta-analysis studies have shown that the accuracy, sensitivity, and specificity of sepsis diagnosis in the early stages of the disease can be dramatically improved, especially when systemic inflammatory response syndrome (SIRS) is combined with CD64+ cells of neutrophils.

[0103] The demonstrated utility of CD64 or a portion thereof as an early marker of sepsis promises the ability to implement innovative treatment strategies (e.g., antibacterial therapy and immunostimulatory and immunosuppressive therapies) that can significantly increase the chances of survival, but such therapies can only be effective if sepsis can be rapidly diagnosed early and the attendant therapies can then be rapidly and continuously implemented thereafter.

[0104] However, currently, flow cytometry is the preferred method for identifying and counting cells presenting specific molecules, including CD64. By detecting and counting individual cells passing through a flow passing through a laser beam, surface immunoglobulins that characterize B cells, T cell receptor-associated molecules known as CD3, CD4 and CD8 coreceptor proteins that distinguish major T cell subsets, and CD64 transmembrane proteins that are upregulated with inflammation, quantitative data can be derived regarding the proportion of cells with different molecules.

[0105] However, the widespread adoption of neutrophil CD64 measurement has been hindered to date because flow cytometry, sophisticated equipment, and well-trained staff with experience are required, despite the need for near-instantaneous diagnosis. CD64-based metrics, such as the neutrophil CD64 (nCD64) ratio, which provides a relative assessment of CD64 expression and neutrophil count, are not considered actionable biomarkers for the development of point-of-care sepsis tests because they rely on flow cytometry to this point and require the identification of specific surface protein expression (CD64) on a particular subset of white blood cells (neutrophils).

[0106] Therefore, there is a need for rapid diagnostic tests for sepsis or severe infections that can assist frontline healthcare providers in providing timely consultation and treatment for the millions of cases of severe bacterial infections in patients admitted to hospitals and intensive care units worldwide.

[0107] A lateral flow assay (LFA) CD64 biomarker (e.g., antigen) configured to examine blood (e.g., whole blood, serum, etc.) provides an alternative tool for the diagnosis of sepsis. As described above, the LFA can be performed on a strip that includes one or more components assembled on a substrate 171. As shown in FIG. 2A, the components of an LFA configured to detect and quantify at least a portion of the CD64 protein in blood and / or other biological fluids can include a sample element 172, a conjugate element 173, a capture element 174, a control element 175, and a wick 176. As shown in FIG. 2B, the components of an LFA configured to detect and quantify at least a portion of the CD64 protein in blood and / or other biological fluids can include a sample element 172, a conjugate element 173, a first capture element 174', a second capture element 174'', a control element 175, and a wick 176. The determination of whether to use one capture element or two or more capture elements corresponding to FIGS. 2A and 2B respectively is determined based on whether one or more portions of CD64 are detected and / or whether a normalization marker (e.g., neutrophil count marker) is detected. In the following description, a more complex scenario of the normalization marker will be described with reference to FIG. 2B, but it can be understood that the same principle can be easily applied to the measurement of at least a portion of CD64 using the LFA of FIG. 2A.

[0108] The substrate 171 can be a backing laminate or backing card configured to provide mechanical support to the components of the LFA, as described above. The substrate 171 can be of any suitable shape, size, and / or configuration, as described above. For example, the substrate 171 can be a rectangular backing card or strip of a certain width and a predetermined length that can provide a sufficient surface area to accommodate the various components of the LFA. The substrate 171 can be made from a semi-rigid polymer designed to provide uniformity and lay-flat characteristics. The substrate 171 can include one or more pressure-sensitive adhesives configured to facilitate the attachment of the various components of the LFA, as further described herein.

[0109] The sample element 172 can be a pad that provides a surface for receiving a sample of blood and / or other biological fluids for analysis and facilitates the transport of the sample in a smooth, continuous, and uniform manner to other components of the lateral flow test strip. The sample element 172 can be of any suitable shape and / or size. In some embodiments, the shape of the sample element 172 can be a rectangular strip configured to adsorb and receive a volume of sample of blood and / or other biological fluids. The sample element 172 can be disposed on the surface of the substrate 171 to provide mechanical support for the LFA. In some embodiments, the sample element 172 can include an adhesive coated on one surface of the sample pad to facilitate attachment to a plastic backing laminate. The shape and dimensions of the sample element 172 can be predetermined such that the sample element can be disposed on the plastic backing laminate. The thickness of the sample element 172 can be selected to facilitate adhesion of the sample element 172 to the plastic backing laminate while maintaining the mechanical structure of the pad. Further, the thickness of the sample element 172 can be selected to accommodate a large volume of blood and / or other biological fluids and prevent oversaturation of the sample on the pad and cross-flow to the plastic backing laminate. The sample element 172 can be made of cellulose, nitrocellulose, glass fiber, and / or any other suitable material.

[0110] In some embodiments, the surface of the sample element 172 can be configured to prepare the sample for subsequent conjugation and capture. For example, understanding that monocytes express CD64 but have no predictive relationship with it, it may be necessary to isolate nCD64 from monocyte CD64 in the sample. In another example, it may be necessary to lyse neutrophils or other cells in the sample to release CD64 from its transmembrane configuration. In yet another example, it may be necessary to cleave the extracellular domain of the neutrophil-derived CD64 protein to enable measurement of this portion.

[0111] For this purpose, various methods are available, for example, to deplete monocytes and / or erythrocytes in the sample and / or to cleave the domain of CD64 for measurement. For example, monocytes in the sample can be depleted by contacting the sample with an anti-CD14 or other related antibody bound to a solid or semi-solid support (i.e., sample element 172). The sample element 172 can be selected for a suitable mesh or pore size and / or can have pads selected by including specific reagents such as antibodies or lectins that bind and retain these components. In another example, erythrocytes in the sample can be depleted by contacting the sample with an anti-glycophorin A antibody bound to a solid or semi-solid support (i.e., sample element 172). In another example, the extracellular domain of the CD64 protein can be cleaved from neutrophils by a protease (e.g., a proteolytic peptidase such as an endopeptidase), and the neutrophils can be captured within the sample element 172.

[0112] For the detection and quantification of CD64 or a portion thereof from a blood and / or other biological fluid sample, the conjugate element 173 of the LFA can be a pad positioned adjacent and downstream of the sample element 172, as shown in FIGS. 2A and 2B. The conjugate element 173 can be of any suitable shape, size, and / or configuration. In some embodiments, the shape of the sample element 172 can be a rectangular strip of a width similar to the width of the sample member 171, disposed on the surface of the substrate 171 to provide mechanical support to the LFA. The conjugate element 173 can include a dry matrix (e.g., a salt-sugar matrix) containing bioactive particles and additives. The bioactive particles contained in the matrix can include specific antibodies and / or affinity reagents (e.g., DNA aptamers, protein binders, etc.) immobilized on or within the conjugate element 173. For example, in some embodiments, the surface of the sample element 172 can be impregnated with an aqueous buffer that provides an environment having a controlled pH. In some embodiments, the surface of the sample element 172 can be impregnated with a buffer solution including, but not limited to, borate buffer solution, phosphate buffered saline (PBS), 2-(N-morpholino)ethanesulfonic acid (MES), tris(hydroxymethyl)aminomethane (TRIS), piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES), 3-(N-morpholino)propanesulfonic acid (MOPS), 2-[4-(2-hydroxyethyl)piperazine-1-yl]ethanesulfonic acid (HEPES), [tris(hydroxymethyl)methylamino]propanesulfonic acid (TAPS), N-cyclohexyl-2-aminoethanesulfonic acid (CHES), and / or N-cyclohexyl-3-aminopropanesulfonic acid (CAPS).

[0113] The dry matrix of sample element 172 can include one or more surfactants used as wetting agents to solubilize polar species present in the sample. For example, in some embodiments, the dry matrix of conjugate element 173 can include nonionic surfactants such as glycidol, tergitol, ethoxylated and alkoxylated fatty acids, ethoxylated amines, alkyl and nonylphenol ethoxylates, ethoxylated sorbitan esters, castor oil ethoxylate, etc. The dry matrix can include one or more biocides configured to promote the extended shelf life of the LFA by inhibiting a wide range of microorganisms. The biocide can be formulated at a low concentration in the dry matrix of conjugate element 173 to minimize and / or avoid potential health hazards, toxicological issues, and disposal problems. For example, in some embodiments, the dry matrix can include other commercially available preservative formulations such as 5-chloro-2-methyl-4-isothiazolin-3-one (CMIT), 2-methyl-4-isothiazolin-3-one (MIT), proprietary glycols, modified alkyl carboxylates, and / or proclin 300 (trademark). In some embodiments, the dry matrix can include one or more surfactants, or any amphiphilic molecule (e.g., tween 20, Triton X, octylthioglucoside, etc.) that can be used for protein solubilization.

[0114] If a normalization marker is detected in addition to CD64 or a portion thereof, the dried matrix of the conjugate element 173 of the LFA in FIG. 2B can include one or more antibodies and / or affinity reagents (177’, 177’’) that conjugate with molecules configured to enable detection. In some embodiments, the dried matrix can include one or more detection antibodies capable of binding to neutrophils or a portion thereof. In one embodiment, the one or more detection antibodies can be, inter alia, antigen-binding fragments that bind to domains of CD64 and / or antigen-binding fragments that bind to a marker of neutrophil count (i.e., a normalization marker). For example, the one or more detection antibodies may be designed to bind to one or more of the extracellular domain, transmembrane domain, and intracellular domain of the CD64 protein, understanding that each domain may present a different antigen. The marker of neutrophil count can be selected to represent the number of neutrophils in the sample and is expected to show a close relationship with CD64 levels in healthy control patients. The neutrophil count marker can be selected from the non-limiting group including neutrophil elastase, lactoferrin, myeloperoxidase, human neutrophil lipocalin, or an equivalent cellular marker whose amount on neutrophils correlates effectively with the neutrophil cell count. In one embodiment, the levels of CD64 and the neutrophil number marker (NNM) can be represented as a detectable signal that can be quantified visually or by an instrument.

[0115] References to the CD64 and NNM biomarkers include their modifications or homolog forms. Modified forms include derivatives, polymorphic variants, truncated forms (truncates), and aggregated or multimeric forms, or forms having extended elements (e.g., amino acid extended elements). In one embodiment, the detection of the total level of CD64 (or CD64 and NNM), the detection of the domains of CD64, and the detection of soluble CD64 or NNM shed from the cell surface (CD64) or the cell surface or interior (NNM) into the plasma fraction are useful for the diagnosis of sepsis.

[0116] The detection antibody can be immobilized on one or more colored particles (e.g., latex having blue color, colloidal gold having red color, and / or any other suitable particles), fluorescent particles, magnetic particles, or any other suitable particles that can be used for capturing and quantifying CD64 and / or NNM in a sample of blood and / or other biological fluids. In some embodiments, the detection antibody can be immobilized on gold nanoparticles. The gold nanoparticles and / or gold nanoshells can be functionalized with an antibody that exhibits specific binding activity to a particular region / domain of the CD64 molecule (e.g., bioconjugation). During bioconjugation, the surface of the gold nanoparticles can be functionalized with the detection antibody using physical methods that rely on physical interactions between the detection antibody and the surface of the gold nanoparticles, such as ionic interactions, hydrophobic interactions, and / or donor bonds. The physical interaction is caused by the spontaneous absorption of the antibody onto the surface of the gold nanoparticles. In the case of ionic interactions, the positively charged groups in the detection antibody are attracted to the negatively charged surface of the gold nanoparticles. Hydrophobic interactions occur between the hydrophobic portion of the detection antibody and the metal surface.

[0117] Alternatively, in some embodiments, the antibody can be conjugated to the gold nanoparticles by a chemical method that involves covalent bonds (e.g., chemisorption via thiol derivatives, bifunctional linkers, and / or adapter molecules). The direct functionalization of gold nanoparticles with thiol derivative groups can be achieved by forming strong bonds on the surface of the particles through a chemical reaction between gold atoms and sulfur atoms. For example, a thiol-functionalized antibody can be directly attached to the gold nanoparticles. However, this approach presents challenges such as potentially compromising the stability of the nanoparticles and the use of reaction conditions that may require stringent conditions.

[0118] The embodiments, implementations, and / or methods described herein can overcome these limitations, for example, by including the use of other groups that can be attached to the surface of gold nanoparticles using a bifunctional linker that provides specific functionalization to the surface of the gold nanoparticles. For example, carboxylated polyethylene glycol (PEG) molecules functionalized with thiol groups (PEG-SH) can be used to functionalize the surface of gold nanoparticles. The PEG molecules functionalized on the gold nanoparticles can also include carboxyl terminal groups. Such carboxyl terminal groups can be modified in a coupling chemical reaction involving water-soluble carbodiimide (EDC) and N-hydroxy-succinimide (NHS) compounds to generate reactive functional groups that bind to the primary amine groups in antibody molecules. The water-soluble carbodiimide reacts with the carboxylic acid moiety in the PEG containing the gold nanoparticles to generate an intermediate active group, which reacts with the N-hydroxy-succinimide compound to form a reactive ester group. The primary amine groups in the antibody react with the ester group formed on the surface of the gold nanoparticles when in direct contact with the antibody. This reaction is designed to generate an amide bond for attaching the antibody to the gold nanoparticles without adding a spacer molecule between the gold nanoparticles and the antibody.

[0119] The capture elements 174’, 174’’ of the LFA for the detection and quantification of CD64 and / or NNM from blood and / or other biological fluid samples can be at least one pad adjacent to and / or downstream of the conjugate element 173, comprising particles or molecules immobilized within or on the capture elements 174’, 174’’ which can be the capture elements. As described above with reference to Figure 2B, the particles or molecules can be configured to bind to the detection antibodies (177’, 177’’) conjugated to the colored particles as described above with reference to the conjugate element 173 when flowing downstream in a volume of blood and / or other biological fluid sample. In some embodiments, the capture elements 174’, 174’’ can include capture antibodies (178’, 178’’) immobilized on and / or chemically bonded to the surface of the capture elements 174’, 174’’. The capture antibodies can be configured to interact with the detection antibodies to capture CD64 and / or NNM bound to the detection antibodies, resulting in a localized accumulation of the detection antibodies and their conjugated colored particles. In some embodiments, the capture antibodies can be adsorbed onto the surface of the capture elements 174’, 174’’.

[0120] The immobilized capture antibodies can be configured to bind to CD64 and / or NNM molecules pre-bound to the detection antibodies (and their conjugated colored particles) at the conjugate element 173. As a result, by exposing the capture elements 174’, 174’’ to a sample of blood and / or other biological fluid containing CD64 and / or NNM that has previously flowed through the conjugate element 173, an accumulation of colored particles associated with the capture antibodies bound to the CD64 and / or NNM molecules present in the sample can be caused. This accumulation of colored particles on the capture elements 174’, 174’’ can be recorded and quantified by one or more optical methods to determine the concentration of CD64 and / or NNM in the sample. For example, in some embodiments, the colored particles accumulated on the capture elements 174’, 174’’ can be determined by a standard lateral flow reader such as the commercially available Leelu reader (LUMOS diagnostics) configured to detect colored particles providing suitable optical sensitivity and dynamic range sufficient to cover a wide range of concentrations.

[0121] The LFA can include a control element 175 for capturing detection antibodies that would otherwise not be captured by the capture elements 174', 174''. In other embodiments, the LFA need not include a control element. The wick 176 of the LFA configured for the detection and quantification of CD64 from a blood and / or other biological fluid sample can be a pad disposed adjacent to and / or downstream of the capture elements 174', 174'' (or the control element 175 if the control element 175 is included). As described with reference to FIG. 2B, the wick 176 can be configured to absorb or wick portions of the sample that are not immobilized within and on the capture elements 174', 174'' (and / or the control element 175 if the control element 175 is included).

[0122] In one embodiment, the detection results of CD64 and / or NNM of the LFA can be one or both of qualitative and quantitative results. For example, if CD64 is detected in the sample and the corresponding capture element visually conveys binding to CD64, a positive determination can be made considering an intensity threshold, a color threshold, or other similar threshold that indicates a level at which sepsis is likely to occur. In another example, binding data from both CD64 and NNM can be considered together to determine a quantitative result. For example, the CD64 data can be normalized by the NNM data.

[0123] In one example, the processing of the binding data and the display of the quantitative result can be performed locally on the LFA or on a fluid transfer and assay system. In another example, the quantitative result or the binding data in the quantitative result can be communicated to an electronic device of the fluid transfer and assay system or to another remotely located electronic device for further analysis. Such further analysis can include computationally intensive analysis of the binding data and potentially more predictive result generation through access to databases, historical data, and sepsis models. Detection is further described in the following paragraphs.

[0124] Generally, determining the levels of CD64 and / or NNM contributes to the establishment of sepsis-related diagnostic rules. Such sepsis-related diagnostic rules can be based on CD64 and / or NNM, and one or more of the biomarkers described herein. The sepsis-related diagnostic rules can be based on the levels or ratios of CD64 and / or NNM relative to a control, since they are related to CD64 alone, or can be based on statistical, variance analysis, and / or machine learning procedures that evaluate the relationship between CD64 and / or NNM and the septic condition.

[0125] For example, such diagnostic rules can be based on the relative levels of CD64 and / or NNM. In one embodiment, when only CD64 is detected and measured, the LFA includes: (a) optionally, contacting the sample with an agent that lyses or solubilizes neutrophils; (b) contacting the sample with a binding agent that specifically binds to the extracellular domain of CD64 in the sample to form a CD64-binding agent complex; and (c) measuring and determining the relative amount of the complex from (b) to obtain a CD64 index that indicates or represents the amount of CD64 in the sample. In one embodiment, the LFA further includes diagnosing that the patient has sepsis or has a risk of developing sepsis if the CD64 index exceeds a threshold above which an excess of neutrophil CD64 is indicated in the sample. The threshold can be adjusted, inter alia, according to the patient's age.

[0126] In one embodiment, when both CD64 and NNM are detected and measured, the LFA includes: (a) optionally, contacting the sample with an agent that lyses or solubilizes neutrophils; (b) contacting the sample with a binder that specifically binds to CD64 in the sample to form a CD64-binder complex and a second binder that specifically binds to a neutrophil marker in the sample to form a neutrophil marker-binder complex; and (c) measuring and determining the relative amounts of each complex from (b) to obtain a corrected CD64 index (the ratio of CD64 to NNM) that indicates or represents the average amount of CD64 per neutrophil in the sample. In some embodiments, the CD64 index from (c) can be corrected for the number of neutrophils in the sample as determined by the amount of neutrophil marker-binder complex measured in (c).

[0127] In one embodiment, the LFA further indicates that a patient has or is at risk of developing sepsis when the CD64 index exceeds a threshold above which an excess of neutrophil CD64 is indicated in the sample. The threshold can be modified, inter alia, according to the age of the patient. In one embodiment, the CD64 index from (b) can be compared to or plotted against the number of neutrophils in the sample as determined by the amount of neutrophil marker-binder complex measured in (c), and the threshold nCD64 index can be corrected for the number of neutrophils in the sample. Since the value of the CD64 index can be a function of the neutrophil count, higher accuracy can be obtained by adjusting the threshold, which was not possible when using flow cytometry alone. The reason is that there is no way to obtain the absolute neutrophil count by flow cytometry without a separate reference method. In one embodiment, a simple algorithm is incorporated or available via the instrument reader to set the threshold for any given value of the neutrophil count. For example, in one embodiment, the threshold is above the 95% confidence interval of the best fit line of the CD64 / NNM ratio, where NNM is neutrophil elastase.

[0128] In one embodiment, when NNM is measured, the ratio of the visual or photometric calculation signal / instrument calculation signal from the CD64 capture element to the visual or photometric calculation signal / instrument calculation signal from the neutrophil marker capture element provides a corrected neutrophil CD64 index indicative of the amount of CD64 per neutrophil in the sample.

[0129] A positive score for sepsis or severe infection or risk according to the present disclosure is determined by comparison of the levels of CD64 and / or NNM and / or CD64 index, enabling rapid administration of antibodies where appropriate. Thus, the present disclosure extends to methods of treatment and prevention, including screening patients according to the immunoassays disclosed herein and administering antibiotics to patients depending on the outcome of the assay. Thus, in another embodiment, the present disclosure teaches the use of the assays and kits and algorithms disclosed herein in the diagnosis and treatment and / or prevention of sepsis or severe infection or risk thereof.

[0130] Furthermore, it should be understood that the rapid test device 170 and / or LFA 170A of FIG. 2B (and / or any other suitable flow-based assay) can be used in conjunction with any of the fluid transfer devices described herein with reference to specific embodiments.

[0131] Aptamer In some implementations, the rapid test device 170 can be an LFA (e.g., LFA 170A or LFA 170B) configured to use aptamers to test for any suitable biomarker associated with sepsis and / or any other infectious condition. For example, the capture elements 174, 174’, 174’’ can be configured to use aptamers to test for procalcitonin, lactate, IL6, CD64, NNM, etc. An aptamer is a single-stranded DNA or RNA molecule that can selectively bind to a corresponding target with high affinity and specificity. These single-stranded molecules consist of a variable region containing 20 to 40 bases at the central end and two constant regions located on both sides thereof and containing binding sites at each end. Aptamers can be folded into secondary and three-dimensional shapes due to intermolecular hybridization. The equilibrium dissociation constant of aptamer-target binding ranges from 1 picomolar (pM) to 1 nanomolar (nM). Aptamers can be generated against desired targets such as toxic small molecules, non-immunogenic targets, or single molecules that do not bind to antibodies, having an affinity similar to that of antibodies for the target molecule. Furthermore, aptamers can be reversibly denatured by heat or chemicals, which is not possible with antibodies.

[0132] Aptamers are similar to antibodies in terms of target recognition and a wide range of applications. However, the use of aptamers presents advantages over the use of antibodies, including ease of control and highly reproducible chemical reactions, production via in vitro processes that rely on chemical reactions, the ability to bind to targets not recognized by antibodies such as ions, small molecules, complex multi-active site molecules, proteins, bacterial cells, viruses, and / or cancer cells, the ability to be amplified in large quantities in a short time by polymerase chain reaction (PCR), ease of modification to introduce functional moieties (such as fluorophores, quenchers, and nanomaterials), stability under harsh conditions, and safety for in vivo use due to their non-immunogenic properties. In some examples, aptamers can improve transport properties, enabling cell-specific targeting and improved tissue penetration.

[0133] Aptamers can be tailored to specific targets obtained through the systematic evolution of ligands by exponential enrichment (SELEX) process. This process involves three main steps: library generation, selection, and amplification. In the first step, a random library is designed and synthesized by combinatorial chemical synthesis techniques to generate oligonucleotides containing variable regions with 20 to 40 bases, flanked on both sides by upstream and downstream primer binding sites at each end. The resulting library is 10 12 ~10 15It may contain individual ssDNA or RNA sequences. In the second step, the target molecule is incubated with the library for several minutes in the presence of a binding buffer. The aptamer binds to the target, forming an aptamer-target complex, and the non-specific sequences remain in the binding buffer. The aptamer-target complex can be recovered and washed several times with a wash buffer. The aptamer can then be separated from the aptamer-target complex by treatment with an elution buffer. The selection step can include a counter-selection procedure in which the target is replaced with itself and nucleic acid sequences that bind to itself are excluded. In the third step, the sequences eluted in the second step are amplified by PCR in the case of DNA or by reverse transcriptase (RT)-PCR in the case of RNA to generate a sub-library for use in the second SELEX process. This procedure can be repeated several times until an aptamer with high specificity for the target is generated.

[0134] When the affinity of the sequences bound to the target is saturated, they are sent to clones and sequences, and subsequently aptamer sequences that bind to the target with high sensitivity and specificity are identified. To improve the separation of unbound sequences from the aptamer-target complex, several techniques can be used. For example, in some examples, aptamer selection can include nitrocellulose membrane filtration-based SELEX, affinity chromatography and magnetic bead-based SELEX, capillary electrophoresis and / or microfluidics-based SELEX. Nitrocellulose membrane filtration-based SELEX uses a nitrocellulose membrane to retain the aptamer-target complex and remove unbound oligonucleotide sequences based on size. Multiple micron-sized pores on the surface of the membrane allow DNA or RNA oligonucleotides to pass through and proteins to be captured on the membrane. The material is then amplified by PCR or RT-PCR for the next production. Affinity chromatography and magnetic bead-based SELEX use agarose beads packed on a column as the stationary phase. Magnetic beads are also used to immobilize the target through physical interaction or chemical reaction between a specific tag and a ligand on the bead. Capillary electrophoresis and microfluidics-based SELEX are used to improve separation speed, resolution, and capacity with minimal sample dilution. In this method, unbound nucleotides are separated from the aptamer-target complex due to the difference in electrophoretic mobility of unbound nucleotides in an electric field. The aptamer can be obtained by the migration speed of a mixture of the target, ligand, or target-ligand complex. Using capillary electrophoresis-based SELEX, aptamers can be selected in several rounds compared to other methods. Microfluidics-based SELEX is a technology that enables automated and miniaturized platforms for aptamer selection on a chip. To perform the selection process automatically, the system includes several modules having micropumps, microvalves, reservoir manifolds, waste chambers, and PCR chambers.Other methods including atomic force microscopy, high-throughput sequencing, graphene oxide, cross-linking by UV, flow cytometry, and surface plasmon resonance (SPR) can be used in connection with the SELEX process. These methods are used to enrich the selection means and improve the efficiency of aptamer selection.

[0135] Aptamer applications include in vivo therapy, molecular biosensors, target capture, drug delivery, new drug development, hazard detection, environmental monitoring, clinical diagnosis, biomarker discovery, and food inspection. Aptamers are used as recognition elements for analytical tools including electrochemical and fluorescent biosensors, colorimetric assays, surface plasmon resonance assays, and amplification techniques.

[0136] Detection In some examples, the rapid test device 170 (e.g., LFA 170A, LFA 170B, and / or any other suitable rapid test device) can be detected and / or determined by a human (e.g., a doctor, a nurse, a technician, etc.) via visual inspection. It can be configured to present test results. For example, a doctor, a nurse, a technician, etc. can visually inspect the capture elements 174, 174’, 174’’ of the LFA 170A or LFA 170B to determine whether a strip is present along the capture elements 174, 174’, 174’’. In addition, the control element 175 of the LFA 170A or LFA 170B can be visually inspected to verify the performance of the test. In some such cases, visual inspection by a human can be relatively simple to implement and may not use additional equipment to provide qualitative results (e.g., positive or negative test results). In other examples, the LFA 170A or LFA 170B can be configured to output raw data (e.g., binding data) and / or test results, and the test results can be received, tested, analyzed, interpreted, etc. by one or more electronic devices (e.g., the electronic device 190 shown in FIG. 1). The binding data can include, for example, electrochemical data, fluorescence data, colorimetric data, optical data, etc. In some examples, a portable strip reader can be used to read, scan, and / or determine a strip along the capture elements 174, 174’, 174’’ and / or the control element 175. The strip reader can include a complementary metal-oxide semiconductor (CMOS) device, a charge-coupled device (CCD), and / or a camera, scanner, reader, etc. that can use any other suitable detection device or camera for detecting the strip. In some implementations, the strip reader can be configured to define data or a digital representation of the test results (strip) that can be qualitative, semi-quantitative, and / or quantitative. For example, the capture element intensity can be proportional to the concentration of the analyte, thereby enabling quantification of the analyte.In some examples, a strip reader may be configured to read, scan, and / or identify the presence and intensity of one or more strips, thereby providing both qualitative and quantitative data. In some implementations, the electronic device 190 may be integrated within / on the rapid inspection device 170, or may be a stand-alone device into which the rapid inspection device 170 and / or one or more cartridges (e.g., one or more parts of the rapid inspection device 170) may be inserted for reading and analysis.

[0137] In some embodiments, the strip reader may be configured to provide qualitative and / or quantitative data as input to the electronic device 190, and the electronic device 190 may analyze, process, and / or otherwise use the data to generate one or more qualitative and / or quantitative test results. The electronic device 190 may be any suitable hardware-based computing device configured to receive, process, define, and / or store data such as, for example, one or more diagnostic test results, test criteria for measuring result data, a predetermined treatment plan and / or a pre-defined treatment plan, a patient profile, a disease profile, etc. Additionally, the electronic device 190 may be configured to transmit and / or receive data via a wired or wireless connection or network. In some embodiments, the electronic device 190 may be, for example, a mobile electronic device (e.g., a smartphone, a tablet, a laptop, and / or any other mobile or wearable device), a personal computer (PC), a workstation, a server device or a distributed network of server devices, a virtual server or machine executed and / or operated as an instance or guest on a physical server or a group of servers, a virtual private server, etc., and / or any other suitable device. In some implementations, the electronic device 190 may be configured to provide a graphical and / or digital representation of the test results generated by the rapid test device 170. Additionally, in some implementations, based on data associated with and / or representing the test results, the electronic device 190 may determine one or more diagnoses, one or more treatment plans, one or more simulations, and / or any other suitable data associated with a body fluid sample, a patient, and / or the treatment of the patient, and may be configured to present it by graphic processing or digital processing.

[0138] In some embodiments, the electronic device 190 can communicate wired or wirelessly with one or more of a plurality of user terminals, such as laptops, desktops, handheld devices, and other user terminals. This communication network can include one or more processors. In one embodiment, the user terminal interacts with one or more processors. In one embodiment, as described above, each processor is implemented using any conventional processing circuit and device or combinations thereof, such as the central processing device (CPU) of a personal computer (PC) or other workstation processor or server, and executes code provided on a hardware computer-readable medium including any conventional memory device to perform any of the methods described herein alone or in combination. In one embodiment, the electronic device 190 and / or one or more processors can be configured to execute at least one algorithm, decision tree, etc. to provide a patient diagnosis based on the input provided to the electronic device 190. In some embodiments, the computational model executed on one or more processors 106 can be configured to predict the sepsis probability (or other prediction).

[0139] For this purpose, in some embodiments, the strip reader can be two or more strip readers configured to provide qualitative and / or quantitative data corresponding to two or more biomarkers as input to the electronic device 190, and the electronic device 190 can analyze, process, and / or otherwise use the data to generate one or more qualitative and / or quantitative test results and / or predictions. For example, the electronic device 190 can receive data corresponding to each of lactate, IL6, PCT, NNM, CD64, etc., and generate one or more qualitative and / or quantitative results and / or predictions therefrom. In one example, the result can include a sepsis score or similar output indicating whether the patient has sepsis or is likely to develop sepsis soon.

[0140] In one embodiment, the collection and analysis of patient data from, for example, a rapid diagnostic test device, and the prediction of patient disease can be performed visually according to visible data generated by the binding of a detection antibody to a capture antibody.

[0141] In one embodiment, the collection and analysis of patient data from, for example, a rapid diagnostic test device, and the prediction of patient disease can be performed by a computational model including, but not limited to, a machine learning model.

[0142] For this purpose, a flowchart for predicting sepsis using a computational model will be described with reference to method 2500 of FIG. 25.

[0143] In some embodiments, at step 2505, qualitative and / or quantitative data corresponding to at least one sepsis-related biomarker can be received by an electronic device from at least one flow-based assay device, such as at least one strip reader. In some variations, the at least one sepsis-related biomarker can include two or more sepsis-related biomarkers. An example is shown in FIG. 1 and described in more detail with reference to FIG. 26. The electronic device (electronic device 190) can be any suitable hardware-based computing device configured to receive, process, define, and / or store data such as, for example, one or more diagnostic test results, test criteria for measuring result data, a predetermined and / or pre-defined treatment plan, a patient profile, a disease profile, etc. In some examples, method 2500 is implemented by electronic device 190, but it should be understood that any computing device configurable to implement the methods and processes herein can be used. For example, as shown in FIG. 25, any device configured to receive data associated with a diagnostic test, an assay, etc. (e.g., rapid test device 170 of FIG. 1), and analyze, process, and / or otherwise use the data to generate one or more qualitative and / or quantitative test results related to the test can be used.

[0144] In step 2510 of method 2500, a computational model can be applied to the received data corresponding to at least one sepsis-related biomarker.

[0145] For example, in some implementations, one or more processors of electronic device 190 (or any other suitable electronic device) can execute a set of instructions associated with one or more computational models based on the application of statistics, analysis of variance, and / or machine learning models and / or procedures. The computational model can be configured to define and / or use the relationships between one or more biomarkers and the disease / health state observed in a control subject to infer one or more relationships, which are then used to predict the state of a patient (e.g., a patient with an unknown state). For example, one or more processors of electronic device 190 and / or any other suitable electronic device can execute and / or use an algorithm that provides and / or outputs a visually detectable score or index of the probability that a patient does not exhibit signs of sepsis or exhibits signs of sepsis or severe infection. In some embodiments, one or more processors of electronic device 190 (or any other suitable electronic device) can execute and / or use an algorithm that implements a multivariate or univariate analysis function.

[0146] In some embodiments, the electronic device 190 and / or one or more of its processors can execute one or more machine learning models that can use, among other things, classifiers such as, for example, nearest neighbor, linear support vector machine, radial basis function support vector machine, decision tree, random forest, AdaBoost, naive Bayes, and / or logistic regression. The desired classifier can be selected through model and parameter optimization. In some applications, the desired classifier can be the classifier having the highest accuracy among all the classifiers tested. In other applications, the desired classifier can be a classifier having the highest positive predictive value, negative predictive value, specificity, selectivity, area under the curve, or any other combination of performance attributes, as defined below. The foregoing provides a subset of suitable approaches for evaluating biomarker data and / or predicting patient situations, but those skilled in the art will readily understand that many other computational models, machine learning concepts, and / or algorithms, including but not limited to artificial neural networks, Bayesian statistics, case-based reasoning, Gaussian process regression, inductive logic programming, learning automata, learning vector quantization, informal fuzzy networks, conditional random fields, genetic algorithms, information theory, support vector machines, averaged one-dependent estimators, group data processing method, instance-based learning, lazy learning, and / or maximum information spanning trees, can be equally used and applied in the methods of the present invention. Further, various forms of enhancement, augmentation, adjustment, optimization, etc. can be applied and / or realized using any combination of the models, processes, algorithms, and / or methods described above.

[0147] In an exemplary machine learning computation set (e.g., the machine learning computation set and / or model described herein with reference to FIG. 25), regardless of which classifier is being used, the original data set can be split in a random fashion into two data sets, a training data set and a test data set, where the training data set includes 80% of the data instances at random (e.g., that an individual patient does not have sepsis [positive] or [negative] at a particular time), and the test data set includes the remaining 20% of the data. Such separation of training data from test data is typical in supervised machine learning applications, such that the model developed during the training phase can be evaluated during the testing phase against data that it has not been previously exposed to. (For example, the test data is equivalent for patients that the model has not been first exposed to, but the model makes predictions about those patients after being exposed to the training data, and then those predictions can be evaluated by comparing them to the test data itself representing those patients).

[0148] When constructing a computational model, the original data set (e.g., training data set, test data set) can have data corresponding to variables, features, and data available to the computational model during runtime (e.g., during implementation in an emergency room or other medical facility when attempting to evaluate a patient's susceptibility to sepsis). In some implementations, it may be desirable for the computational model to be based on the fewest number of variables that give an accurate (or useful) result. This ensures that the computational model can be implemented anywhere at the point of care.

[0149] In some embodiments, the training dataset can include variables measurable at the point of care and / or variables obtainable from medical records. Variables measurable at the point of care can include, for example, one or more biomarkers (e.g., lactate, IL6, CD64, PCT, and / or NNM as described herein), heart rate, blood pressure, white blood cell count, respiratory rate, body temperature, etc. (commonly referred to as “point of care” measurements, variables, metrics, and / or data). In some implementations, data corresponding to point of care variables can be obtained, for example, from the electronic health records of previous patients. Variables obtained from medical records can include medical history data, previous diagnoses, treatment plans and medications, tests and test results, immunization details and dates, medical images (e.g., radiographic images), etc. (commonly referred to as “history” measurements, variables, metrics, and / or data). Data corresponding to medical history variables can similarly be obtained from the electronic health records of previous patients. In some embodiments, the training dataset can include variables measurable at the point of care and variables obtained from medical records, thereby providing past context to point of care measurements.

[0150] In some embodiments, a patient population dataset can be provided to a computer system, and the system selects a plurality of subsets from the patient population dataset. Each subset is used by a machine learning algorithm applied by the system to each respective subset to train a new prediction model, and based on this prediction model, the onset of a disease, e.g., sepsis, in a patient is predicted. Thus, for each selected subset, a respective prediction model can be trained, and each of the trained prediction models is then applied to the data of individual patients regarding a particular group of characteristics of the subset on which that respective prediction model was trained. This ensures that the prediction model (or computational model, or machine learning model) is based on data that the patient and / or the immediate patient care environment can provide.

[0151] When returning to the computational model, for each of these classifiers, the model parameters that determine their prediction models can be calculated based on the training dataset. For logistic regression, the parameters for each resulting model are the sum of a single bias value and one coefficient for each data feature in the model. The data features are a type of measurement (e.g., systolic blood pressure measurement). The linear combination of the coefficients and the normalized data features (patient data), along with the bias value, generates a prediction.

[0152] Next, each classifier model is used with its own respective set of parameters obtained from the training dataset (as described above), evaluated on the test dataset, and can yield prediction results expressed in terms of accuracy, positive predictive value (PPV), sensitivity, specificity, negative predictive value (NPV), and area under the curve (AUC).

[0153] Returning to method 2500, at step 2515, a sepsis probability score can be generated based on the output of the applied computational model.

[0154] In some embodiments, a probability greater than 50% (1 / 2) results in a prediction that the patient will have sepsis at the corresponding future time point, and a probability of 50% (1 / 2) or less is a prediction of no sepsis. As will be understood by those skilled in the art, it is straightforward to apply more sophisticated processing of this probability to assign finer priorities and / or categories to the likelihood and severity of a situation. For example, the above probability can be used directly as a measure of the predicted probability of developing sepsis, in which case, rather than a binary prediction of which patients will or will not develop sepsis, the probability can be mapped to categories such as "high likelihood of developing sepsis", "high probability of developing sepsis", "possible to develop sepsis", "low likelihood of developing sepsis", and "very low likelihood of developing sepsis". These finer priorities and / or categories can be particularly useful for a hospital when acting based on the prediction.

[0155] In optional step 2520 of method 2500, based on the sepsis probability score generated in step 2515, a healthcare provider can be alerted to initiate a corresponding treatment when the sepsis probability score exceeds a predetermined sepsis-related threshold. For example, as described above, a sepsis probability score greater than 50% can be an indicator that a sepsis treatment plan as described above should be initiated.

[0156] As described above, in some implementations, the transfer device 105 can be configured to transfer a first quantity of body fluid to the rapid test device 170 and transfer at least a portion of a second quantity of body fluid to one or more of the optional fluid collection devices 195. For example, the second portion or sampling portion of the transfer device 105 can include an outlet or port that can enable the second quantity of body fluid to be transferred from the second portion or sampling portion of the transfer device 105, and / or can be in fluid communication with the outlet or port. In some examples, one or more optional fluid collection devices 195 can be physically and / or fluidly coupled to the transfer device 105 (e.g., via an outlet or port) to receive at least a portion of the second quantity of body fluid.

[0157] In some embodiments, the optional fluid collection device 195 can be any suitable device for at least temporarily containing a body fluid. For example, the fluid collection device 195 can include, but is not limited to, any suitable container, receptacle, reservoir, bottle, adapter, dish, vial, syringe, device, diagnostic and / or testing machine, etc. In some embodiments, the fluid collection device can be substantially similar to or the same as a known sample container such as, for example, a Vacutainer® (manufactured by Becton Dickinson and Company (BD)), a BacT / ALERT® SN or BacT / ALERT® FA (manufactured by Biomerieux, Inc.), and / or any suitable reservoir, vial, microvial, microliter vial, nanoliter vial, container, microcontainer, nanocontainer, and / or the like. In some embodiments, the fluid collection device can be substantially similar to or the same as any of the sample reservoirs described in U.S. Patent No. 8,197,420, entitled "Systems and Methods for Parenterally Procuring Bodily-Fluid Samples with Reduced Contamination," filed December 13, 2007 (the " '420 Patent"), the disclosure of which is hereby incorporated by reference in its entirety.

[0158] In some embodiments, the fluid collection device 195 can drain its contents prior to receiving a sample volume of body fluid. For example, in some embodiments, the fluid collection device 195 or reservoir can be configured to define or create vacuum, suction, and / or negative pressure conditions, such as a vacuum-based collection tube (e.g., Vacutainer®), a syringe, etc. In some implementations, the fluid collection device 195 can be physically and / or fluidly coupled to the transfer device 105 (e.g., an outlet or port), whereby, as described in more detail herein with reference to certain embodiments, the negative pressure conditions within the fluid collection device 195 facilitate the withdrawal of body fluid from the patient and the drawing of the body fluid into one or more portions of the transfer device 105, or through the transfer device 105.

[0159] In some embodiments, the fluid collection device 195 can contain any suitable additive, culture medium, substance, enzyme, oil, fluid, etc. For example, the fluid collection device 195 can be, for example, a sample or culture bottle containing an aerobic or anaerobic culture medium. The sample or culture bottle can be configured to receive a body fluid sample, which can then be tested (e.g., after culturing using in vitro diagnostic (IVD) tests and / or any other suitable tests) for the presence of, for example, gram-positive bacteria, gram-negative bacteria, yeast, fungi, and / or any other organism. In some examples, if such a test of the culture medium results in a positive result, the culture medium can then be tested using a nucleic acid-based system (e.g., a PCR-based system, hybridization probe, nucleic acid amplification test (NAAT), etc.) to identify the specific organism. In some embodiments, the sample reservoir can contain, for example, any suitable additive, etc., in addition to or instead of the culture medium. Such additives can include, for example, heparin, citrate, ethylenediaminetetraacetic acid (EDTA), oxalate, sodium polyanethol sulfonate (SPS), etc. In some embodiments, the fluid collection device 195 can contain any suitable additive or culture medium, and can be evacuated and / or air can be removed from the fluid collection device 195 in other ways.

[0160] The "culture medium" has been described above as a substance configured to react with organisms (e.g., microorganisms such as bacteria) in body fluids, and the "additive" has been described above as a substance configured to react with a part of body fluids (e.g., blood constituent cells, serum, synovial fluid, etc.). However, it should be understood that the sample reservoir can contain any suitable substance, liquid, solid, powder, lyophilized compound, gas, etc. Further, when referring to the "additive" in the sample reservoir, the additive can be a culture medium such as an aerobic culture medium and / or an anaerobic culture medium contained in the culture bottle, an additive contained in the culture bottle and / or other suitable reservoir described above, and / or any other suitable substance or combination of substances. That is, the embodiments described herein can be used with any suitable fluid reservoir containing any suitable substance or combination of substances, etc.

[0161] In some implementations, a second quantity of body fluid contained within the second portion or sampling portion of the transfer device 105 and / or within optional one or more fluid collection devices 195 can be used as a biological sample in one or more tests, assays, and / or diagnostic methods. In some examples, by isolating the first quantity of body fluid from the second quantity of body fluid, contaminants within the first quantity of body fluid and / or within the isolated portion of the transfer device 105 can be isolated. By isolating, the second quantity of body fluid can be kept substantially free of contaminants. Thus, the second portion or quantity of body fluid, which may be relatively sensitive to contaminants (e.g., can result in poor results due to the presence of contaminants), can be used in one or more tests such as a blood culture test. In this way, the system 100 can be configured to obtain a first quantity of body fluid that can be used in tests having a relatively low sensitivity to contamination and a second quantity of body fluid that can be used in tests having a relatively high sensitivity to contamination. In some examples, the test of the first quantity of body fluid can provide relatively rapid initial results that can inform one or more treatment options, while the test of the second quantity of body fluid can provide more detailed test results that typically take longer to conduct. Thus, in the case of time-critical disease conditions (e.g., sepsis), the initial results from the test of the first quantity of body fluid can enable a physician or clinician to provide rapid initial treatment while a more detailed test of the second quantity of body fluid is being performed.

[0162] FIG. 3 is a schematic diagram of a fluid transfer and assay system 200 according to one embodiment. The fluid transfer and assay system 200 (also referred to herein as the “system”) can include at least a fluid transfer device 205 and a rapid diagnostic test device 270. Additionally, the system 200 can include at least one fluid collection device 295 that can be physically and / or fluidly coupled to the fluid transfer device 205.

[0163] The fluid control device 205 (also referred to herein as the "transfer device") can be of any suitable shape, size, and / or configuration. In some implementations, the transfer device 205 can be configured to draw body fluid (e.g., blood) from a patient into and / or through the transfer device 205. Additionally, the transfer device 205 can be configured to transfer at least a portion of the drawn body fluid to one or more other devices, reservoirs, containers, vials, machines, tests, assays, etc., such as the rapid diagnostic test device 270 and / or one or more sample reservoirs 295.

[0164] The transfer device 205 includes a housing 210 and a catheter 250. The housing 210 of the device 205 can be of any suitable shape, size, and / or configuration. For example, in some embodiments, the housing 210 can have a size that is at least partially based on an initial amount or volume of body fluid that is configured to be transferred within and / or isolated within a portion of the housing 210. In some embodiments, the housing 210 can have a size and / or shape that is configured to improve the ergonomic performance and / or ease of use associated with the device 205. Further, in some embodiments, one or more portions of the housing 210 can be formed from a relatively transparent material that is configured to allow a user to visually inspect and / or verify the flow of body fluid through at least a portion of the housing 210.

[0165] The housing 210 has and / or forms an inlet 212 and an outlet 213, and defines at least one fluid flow path 215 therebetween. The inlet 212 can be any suitable inlet, opening, port, stopcock, lock (e.g., luer lock), seal, coupler, valve (e.g., one-way valve, check valve, duckbill valve, umbrella valve, etc.), tube, conduit, etc. The inlet 212 is configured to fluidly couple the housing 210 to a body fluid source (e.g., a patient). For example, the inlet 212 can be connected to an intravascular device (e.g., a winged needle, an intravenous (IV) catheter, a peripherally inserted central catheter (PICC), a midline, an intermediate intravascular device, etc.) configured to be placed percutaneously within the patient. Thus, fluid can be transferred between the housing 210 and the patient via the inlet 212 and any intravascular device connected between the housing 210 and the patient. More specifically, the transfer device 205 can be configured to transfer body fluid from the patient and / or any other body fluid source, through the inlet 212 (and / or any intravascular device connected thereto), and into the housing 210 via the inlet 212, as described in more detail herein.

[0166] As shown in FIG. 3, the housing 210 defines one or more fluid flow paths 215 between the inlet 212 and the outlet 213. As described in more detail herein, the transfer device 205 and / or the housing 210 can be configured to transition between any number of states, operating modes, and / or configurations to selectively control the flow of body fluid through the one or more fluid flow paths 215. Further, the transfer device 205 and / or the housing 210 can be configured to transition automatically (e.g., based on a pressure differential, time, electronic control, membrane saturation, absorbent material and / or barrier material, etc.) or via an intervention (e.g., user intervention, mechanical intervention, etc.).

[0167] Outlet 213 is in fluid communication with one or more fluid flow paths 215 and is configured to selectively receive the flow of bodily fluid from inlet 212 (via fluid flow path 215). Outlet 213 can be any suitable outlet, opening, port, stopcock, lock, seal, coupler, valve, tube, conduit, etc. that is configured to be physically and / or fluidly coupled to any suitable device that is coupled to outlet 213, such as, for example, fluid collection device 295 (e.g., a fluid or sample reservoir, syringe, vacuum container, culture bottle, etc.). In some embodiments, outlet 213 can be formed monolithically with fluid collection device 295. In other embodiments, outlet 213 can be at least temporarily coupled to fluid collection device 295 via an adhesive, interference fit, mechanical fastener, screw connection, piercing or puncture array configuration, any number of mating recesses, and / or other suitable joints or combinations thereof. For example, in some embodiments, outlet 213 can include and / or be coupled to a fluid transfer adapter, such as the fluid transfer adapter described in U.S. Patent No. 10,123,783, entitled "Apparatus and Methods for Disinfection of a Specimen Container," filed Mar. 2, 2015 ("the '783 patent"), and / or can be coupled to a fluid transfer device, such as the fluid transfer device described in U.S. Patent Application Publication No. 2015 / 0342510, entitled "Sterile Bodily-Fluid Collection Device and Methods," filed Jun. 2, 2015 ("the '510 application publication"), the disclosures of each being incorporated herein by reference in their entireties. In such embodiments, the fluid transfer adapter can be coupled to and / or receive a portion of fluid collection device 295 and can establish fluid communication between outlet 213 and fluid collection device 295. In still other embodiments, outlet 213 can be operably coupled to fluid collection device 295 via an intervening structure (not shown in FIG. 3), such as a sterilization tube.

[0168] In some embodiments, the arrangement of the outlets 213 can be such that the outlets 213 are physically and / or fluidly sealed before being connected to the collection device 295. In some embodiments, the outlet 213 can transition from a sealed configuration to an unsealed configuration in response to being connected to the collection device 295 and / or in response to a negative pressure differential between the environment within the outlet 213 and / or the housing 210 and the environment within the fluid collection device 295.

[0169] The fluid collection device 295 can be any suitable device for at least temporarily containing body fluid, such as any of the devices described in detail above with reference to the fluid collection device 195 (e.g., a vacuum container, a sample reservoir, a syringe, a culture bottle, etc.). In some embodiments, the fluid collection device 295 can be a sample reservoir that includes a vacuum seal to maintain negative pressure conditions (vacuum conditions) within the sample reservoir, and thus can facilitate drawing body fluid from the patient into the sample reservoir through the transfer device 205 using a vacuum or suction force. In embodiments where the fluid collection device 295 is a vacuum container or the like, the user can connect the fluid collection device 295 to the outlet 213 to initiate the flow of body fluid from the patient to the device 205, and as further described in more detail herein, the first or initial portion of the flow of body fluid can be transferred and / or isolated to, for example, the rapid diagnostic test device 270, and the second or subsequent portion of the flow of body fluid can be diverted away from the rapid diagnostic test device 270 and to the fluid collection device 295 (e.g., via the outlet 213) and / or otherwise shunted.

[0170] The actuator 250 of device 205 is at least partially disposed within the housing 210 and is configured to control, direct, and / or otherwise facilitate a selective flow of fluid through at least a portion of the housing 210 and / or at least a portion of one or more fluid flow paths 215. The actuator 250 can be of any suitable shape, size, and / or configuration. In some embodiments, the actuator 250 can be a member or device configured to transition between two or more states (e.g., at least a first state and a second state). For example, the actuator 250 can be a valve, a plunger, a seal, a membrane, a bladder, a flap, a plate, a rod, a switch, etc. The actuator 250 can be actuated and / or transitioned between any number of states (e.g., at least a first state and a second state) in any suitable manner. For example, transitioning the actuator 250 can include actuating, pushing, moving, translating, rotating, switching, sliding, opening, closing, and / or otherwise reconfiguring the actuator 250.

[0171] In some implementations, the actuator 250 can be configured to transition between at least a first state and a second state in response to manual actuation by a user (e.g., manually applying force to a button, slider, plunger, switch, valve, rotating member, conduit, etc.). In other implementations, the actuator 250 can be configured to automatically transition between at least a first state and a second state in response to a pressure difference (or lack thereof), a change in potential or kinetic energy, a change in composition or configuration (e.g., a part of the actuator can at least partially dissolve or deform), etc. In yet other implementations, the actuator 250 can be mechanically and / or electrically actuated or transitioned based on a predetermined time, the volume of received body fluid, the volumetric flow rate of the body fluid flow, the flow velocity of the body fluid flow, etc. (e.g., can be actuated or transitioned via a motor, a spring release mechanism, etc.). Examples of actuators and / or ways in which the actuator can transition are provided, but it should be understood that they are not limiting and are presented only by way of example.

[0172] In the embodiment shown in FIG. 3, the actuator 250 can be configured to selectively establish fluid communication between the inlet 212 and the rapid diagnostic test device 270 when in the first state, and to selectively establish fluid communication between the inlet 212 and the outlet 213 when in the second state. When in the first state, the actuator 250 can be configured to allow body fluid to flow from the inlet 212, through at least a portion of the fluid flow path 215, and into the rapid diagnostic test device 270 or to flow into the rapid diagnostic test device 270. In some embodiments, the actuator 250 can be configured to isolate, separate, and / or otherwise prevent fluid communication between the outlet 213 and the inlet 212, at least a portion of the fluid flow path 215, and / or the rapid diagnostic test device 270. When in the second state, the actuator 250 can be configured to allow a subsequent volume of body fluid (e.g., a volume of body fluid after an initial volume of body fluid) to be transferred from the inlet 212, through at least a portion of the fluid flow path 215, to the outlet 213 (and / or a fluid collection device 295 fluidly coupled to the outlet 213), as described in more detail herein. Additionally, when in the second state, the actuator 250 can be configured to isolate, separate, and / or otherwise prevent fluid communication between the rapid diagnostic test device 270 and the inlet 212, the outlet 213, and / or at least a portion of the fluid flow path 215, as described in more detail herein.

[0173] The rapid diagnostic test device 270 (also referred to herein as the "rapid test device" or simply the "test device") can be of any suitable shape, size, and / or configuration. In some embodiments, the rapid test device 270 can be removably coupled to the transfer device 205 or any suitable portion thereof (e.g., an inlet portion, an outlet portion, an isolation portion, a sampling portion, and / or any other suitable portion). In other embodiments, the rapid test device 270 can be integrated into the transfer device 205. For example, the transfer device 205 and the rapid test device 270 can be formed integrally or monolithically and / or integrated in other ways. In still other embodiments, the transfer device 205 can include a port, an adapter, and / or a receiving portion to which the rapid test device 270 can be coupled or into which the rapid test device 270 can be inserted to establish fluid communication between the transfer device 205 and the rapid test device 270 and / or can be formed to include such. In some such embodiments, coupling the rapid test device 270 to the transfer device 205 can act to transition one or more flow controllers, valves, partitions, ports, seals, etc. from a closed or sealed state to an open state to enable fluid communication between the transfer device 205 and the test device 270.

[0174] In some implementations, the rapid test device 270 can be configured to receive a first volume of body fluid from the transfer device 205 and perform one or more tests, assays, and / or diagnostic methods using the first volume of body fluid. The rapid test device 270 can be any suitable test device. For example, the rapid test device 270 can be an LFA or the like, as described in detail above with reference to the LFA170A or LFA170B shown in FIGS. 2A and 2B, respectively. In some implementations, the test device 270 can be an LFA configured to test for the presence of a specific analyte or biomarker that can provide information used to diagnose a patient condition, such as sepsis and / or any other disease state. For example, the LFA can be configured to test for lactate, IL6, PCT, CD64, and / or NNM biomarkers that can be indicators of sepsis. In other embodiments, the test device can be an LFA configured to test for any of the target analytes and / or biomarkers described above with reference to the LFA170A or LFA170B shown in FIGS. 2A and 2B, respectively.

[0175] In some examples, the rapid test device 270 can be configured to output test results associated with testing the volume of body fluid transferred from the transfer device 205 while the transfer device 205 and / or the actuator 250 is in a first state. The test results (indicated by the arrow labeled "output" in FIG. 3) can be detected and / or determined by a human via visual inspection and / or by one or more electronic devices (e.g., the electronic device 290). In some examples, the test results output by the rapid test device 270 can be qualitative, semi-quantitative, and / or quantitative. Thus, the rapid test device 270 may be structurally and / or functionally similar or identical to the rapid test device 170 described in detail above, and thus will not be described in further detail herein.

[0176] As described above, system 200 can be used to obtain from a patient one or more volumes of body fluid that can be used in one or more tests, assays, and / or diagnostic methods. For example, in some instances, a user such as a physician, internist, nurse, phlebotomist, technician, etc. can operate device 205 to establish fluid communication between inlet 212 and a body fluid source (e.g., a patient's vein, cerebral spinal fluid (CSF) from the spinal cavity, urine collection, etc.). As a specific example, in some instances, inlet 212 can be connected to and / or include a needle or the like that is operable to pierce the patient's skin and insert at least a portion of the needle into the patient's vein, thereby positioning inlet 212 to fluidly communicate with a body fluid source (e.g., a vein, an IV catheter, a PICC, etc.).

[0177] In some instances, actuator 250 can be in a first state when inlet 212 is positioned in fluid communication with a body fluid source (e.g., a portion of a patient), thereby establishing fluid communication between at least a portion of fluid flow path 215 between inlet 212 and rapid test device 270 (and / or a portion of device 205 to which rapid test device 270 is connected). Accordingly, transfer device 205 can be configured to transfer an initial volume of body fluid from a body fluid source (e.g., a patient) to rapid test device 270. In some implementations, the initial volume of body fluid can flow passively (e.g., without user intervention and / or transition of one or more components) to rapid test device 270 in response to a positive pressure associated with the patient's vasculature and / or in response to any of the fluid transfer methods described in U.S. Patent Application Publication No. 2018 / 0353117, titled "Fluid Control Devices and Methods of Using the Same," filed on June 11, 2018 (the "'117 application publication"), the disclosure of which is incorporated herein by reference in its entirety.

[0178] In other embodiments, the transfer device 205 and / or a portion thereof can be configured to create a negative pressure differential (e.g., partial vacuum, suction, etc.) in at least a portion of the fluid flow path 215 that can initiate and / or maintain a flow of an initial volume of bodily fluid from the bodily fluid source to the rapid test device 270. For example, in some examples, the actuator 250 can be stored in a third state (e.g., a storage state) prior to use and transitioned from the storage state to the first state to initiate a flow of an initial volume of bodily fluid. In such examples, transitioning the actuator 250 can create a negative pressure that can draw bodily fluid from the inlet 212 into the rapid test device 270. In some such embodiments, the actuator 250 is in the same and / or substantially the same manner as described in any of U.S. Patent No. 8,535,241, entitled "Fluid Diversion Mechanism for Bodily-Fluid Sampling," filed October 22, 2012 (the " '241 Patent"), U.S. Patent No. 9,060,724, entitled "Fluid Diversion Mechanism for Bodily-Fluid Sampling," filed May 29, 2013 (the " '724 Patent"), U.S. Patent No. 9,155,495, entitled "Syringe-Based Fluid Diversion Mechanism for Bodily-Fluid Sampling," filed December 2, 2013 (the " '495 Patent"), U.S. Patent No. 11,234,626, entitled "Devices and Methods for Syringe Based Fluid Transfer for Bodily-Fluid Sampling," filed June 23, 2016 (the " '626 Patent"), and / or U.S. Patent Application Publication No. 2020 / 0253524, entitled "Devices and Methods for Bodily Fluid Collection and Distribution," filed February 7, 2020 (the " '524 Publication"), and can be transitioned to create a negative pressure differential. The disclosure of each of these patents and patent applications is hereby incorporated by reference in its entirety.In yet other implementations, the initial volume of body fluid can flow to the rapid test device 270 in response to the negative pressure differential generated by the fluid collection device 295, as described in more detail herein with reference to other embodiments.

[0179] The initial volume of body fluid can be any suitable volume of body fluid, such as any of the volumes or amounts described above. For example, in some instances, the transfer device 205 can remain in a first state or configuration until a predetermined and / or desired volume (e.g., the initial volume) of body fluid has been transferred to the rapid test device 270. In some embodiments, the initial volume can be associated with and / or at least partially based on a desired volume sufficient for the rapid test device 270 to perform one or more tests or assays. In other embodiments, the initial volume of body fluid can be associated with and / or at least partially based on an amount or volume of body fluid that is greater than the volume associated with the fluid flow path defined between the body fluid source and the rapid test device 270. In yet other embodiments, the transfer device 205 can be configured to transfer a certain flow rate of body fluid (e.g., the initial volume) to the rapid test device 270 until the pressure differential between the rapid test device 270 and the inlet 212 or body fluid source has been brought to a substantially equilibrium state and / or otherwise reduced below a desired threshold.

[0180] In some embodiments, the rapid test device 270 can initiate the testing and / or analysis of an initial volume of a body fluid, such as when the initial volume is transferred to, for example, a sample element (such as sample element 171). In some examples, the rapid test device 270 can be configured to provide one or more solutions, buffers, mixtures, additives, etc. that can be mixed or combined with the initial volume. In this way, the initial volume of the body fluid (whether alone or mixed with additional components) can flow through the rapid test device 270 (such as an LFA as described above with reference to FIGS. 2A or 2B), and the rapid test device 270 can then perform one or more tests or assays on the initial volume. For example, in some examples, the rapid test device 270 can be an LFA configured to test for the presence of lactate, IL6, PCT, CD64, and / or NNM, as described in detail above. Further, upon completion of the test or assay, the rapid test device 270 can be configured to output the test result, which can be detected and / or determined by a human and / or one or more electronic devices, as described in detail above.

[0181] After the initial volume of the body fluid has been transferred and / or diverted into the rapid test device 270, the transfer device 205 can transition from a first state or configuration to a second state or configuration. For example, in some embodiments, the actuator 250 can transition from a first state of the actuator 250 to its first state when the initial volume of the body fluid is transferred into the rapid test device 270, thereby placing the transfer device 205 in its second state. Further, in some embodiments, the arrangement configuration of the transfer device 205 can be such that the transfer device 205 cannot transition to its second state before collecting the initial volume within the rapid test device 270.

[0182] In some embodiments, the arrangement of the transfer device 205, the actuator 250, and / or the rapid test device 270 can be such that in response to receiving the initial volume, the flow of body fluid into the rapid test device 270 substantially stops or slows down. In some examples, the user can visually inspect a portion of the device 205 and / or the housing 210 to determine that the initial volume of body fluid has been placed within the rapid test device 270 and / or that the flow of body fluid into the rapid test device 270 has slowed or substantially stopped. In some embodiments, the user can apply a force to the actuator 250 and / or otherwise activate the actuator 250 to transition the actuator 250 from its first state to its second state. In other embodiments, the actuator 250 can be automatically transitioned (e.g., without user intervention). Further, in some implementations, the device 205 and / or the actuator 250 can transition from a first state to a second state while the rapid test device 270 is performing an assay on the initial volume of body fluid. Stated another way, the rapid test device 270 can perform an assay on the initial volume of body fluid while the device 205 is being used to transfer one or more subsequent volumes of body fluid (e.g., in one or more parallel processes, etc.).

[0183] In some embodiments, transitioning the actuator 250 from its first state to its second state (e.g., placing the transfer device 205 in its second state or configuration) can isolate, separate, and / or retain an initial volume of body fluid within the rapid test device 270. Said another way, the actuator 250 can isolate and / or separate the rapid test device 270 from one or more portions of the inlet 212, outlet 213, and fluid flow path 215. As described in more detail herein, in some examples, contaminants such as microorganisms present on the skin removed during a venipuncture event, other external contamination sources, and contaminants such as catheter and PICC line colonization used to collect a sample can be mixed into and / or contained within the initial volume of body fluid. Thus, such contaminants are isolated within the initial volume. Further, the arrangement configuration of the rapid test device 270 can be such that the tests and / or assays performed by the rapid test device 270 are not affected by such contamination, which means that the accuracy of the test results output by the rapid test device 270 is not affected by such contamination, as detailed above.

[0184] In addition to isolating the rapid test device 270 from at least a portion of the inlet 212, outlet 213, and fluid flow path 215, placing the actuator 250 in its second state also establishes fluid communication between the inlet 212 and the outlet 213 through at least a portion of the fluid flow path 215. For example, in some embodiments, transitioning the actuator 250 from its first state to its second state can, for example, open or close a port or valve, move one or more seals, move or remove one or more obstacles, define one or more portions of the flow path, and the like.

[0185] In some implementations, the fluid collection device 295 can be fluidly connected to the outlet 213 at any time before and / or simultaneously with the actuator 250 transitioning from the first state to the second state. As described above, the fluid collection device 295 can be any suitable reservoir, container, and / or device configured to receive a volume of body fluid. For example, the fluid collection device 295 can be a vacuum reservoir or container that defines a negative pressure and / or can be a syringe that is operated to generate a negative pressure. In some examples, by connecting the outlet 213 to the fluid collection device 295, at least a portion of the fluid flow path 215 is selectively exposed to the negative pressure and / or suction force within the fluid collection device 295. Thus, in response to the negative pressure and / or suction force, one or more subsequent volumes of body fluid can flow from the inlet 212, through at least a portion of the fluid flow path 215, through the outlet 213, and into the fluid collection device 295. As described above, by sequestering an initial volume of body fluid (e.g., within the rapid test device 270) prior to collecting or acquiring one or more subsequent volumes of body fluid, the amount of contaminants in the one or more subsequent volumes is reduced and / or substantially eliminated. Thus, subsequent volumes of body fluid, which may be relatively sensitive to contaminants (e.g., can result in poor results due to the presence of contaminants), can be used in one or more tests such as blood culture tests. In this way, the system 200 can be configured to acquire an initial volume of body fluid that can be used in tests with a relatively low sensitivity to contamination and a subsequent volume of body fluid that can be used in tests with a relatively high sensitivity to contamination. In some examples, testing of the initial volume of body fluid (e.g., by the rapid test device 270) can provide relatively rapid initial results that can inform one or more treatment options, while testing of the subsequent volume of body fluid can typically provide more detailed test results that take longer to develop.

[0186] Figure 4 is a schematic diagram of a fluid transfer and assay system 300 according to one embodiment. The fluid transfer and assay system 300 (also referred to herein as the "system") can include at least a fluid transfer device 305 and a rapid diagnostic test device 370. In some implementations, the system 300 can include at least one fluid collection device 395 that can be physically and / or fluidly coupled to the fluid transfer device 305. Some portions and / or aspects of the fluid transfer device 305, the rapid diagnostic test device 370, and / or the fluid collection device 395 are each similar to and / or can be substantially the same as the fluid transfer devices 105 and / or 205, the rapid diagnostic test devices 170 (and / or LFA 170A, 170B) and / or 270, and / or the fluid collection devices 195 and / or 295 described in detail above with reference to FIG. 3. Accordingly, some such portions and / or aspects are not described in further detail herein.

[0187] The fluid transfer device 305 (also referred to herein as the "transfer device") can be of any suitable shape, size, and / or configuration. In some implementations, the transfer device 305 can be configured to draw a body fluid (e.g., blood) from a patient into the transfer device 305 and / or through the transfer device 305. Additionally, the transfer device 305 can be configured to transfer at least a portion of the drawn body fluid to one or more other devices, reservoirs, containers, vials, machines, tests, assays, etc., such as the rapid diagnostic test device 370 and / or one or more fluid collection devices 395.

[0188] The transfer device 305 includes a housing 310, a flow computer 340, and an actuator 350. The housing 310 of the device 305 can have any suitable shape, size, and / or configuration. For example, in some embodiments, the housing 310 can be the same as or substantially the same as the housing 210 described above with reference to FIG. 3. Specifically, the housing 310 has and / or forms an inlet 312 and an outlet 313, and defines at least one fluid flow path 315 between the inlet 312 and the outlet 313. The inlet 312 can be any suitable inlet or port and can be configured to establish fluid communication between the housing 310 and a body fluid source (e.g., a patient). The outlet 313 can be any suitable outlet or port and can be configured to establish fluid communication between the housing 310 and the fluid collection device 395. Further, the fluid collection device 395 can be the same as or substantially the same as the fluid collection device 295 and thus will not be described in further detail herein. The one or more fluid flow paths 315 defined by the housing 310 extend between the inlet 312 and the outlet 313 and can selectively establish fluid communication between the inlet 312 and the outlet 313 as described in further detail herein.

[0189] However, the housing 310 can be different from the housing 210 by including, forming, and / or being connected to an isolation chamber 330. As described in further detail herein, the isolation chamber 330 is selectively in fluid communication with the fluid flow path 315. In addition, the isolation chamber 330 includes, is connected to, and / or is otherwise in fluid communication with a rapid diagnostic test device 370. The isolation chamber 330 can be configured to (1) receive a certain flow rate and / or volume of body fluid from the inlet 312, (2) isolate (e.g., separate, divide, contain, hold, isolate, etc.) at least a portion of the certain flow rate and / or volume of body fluid within the isolation chamber 330, and (3) transfer at least a portion of the certain flow rate and / or volume of body fluid into the rapid diagnostic test device.

[0190] The isolation chamber 330 can have any suitable array configuration, such as the array configuration described herein with respect to certain embodiments. For example, in some embodiments, the isolation portion 330 can be at least partially formed by the housing 310. In other embodiments, the isolation portion 330 can be a reservoir placed within and / or disposed within a portion of the housing 310. In other embodiments, the isolation chamber 330 can be formed and / or defined by a portion of the fluid flow path 315. That is, the housing 310 can define one or more lumens and / or can include one or more lumen-defining devices configured to receive an initial fluid or volume of body fluid from the inlet 312, thereby forming the isolation chamber 330 and / or functioning as the isolation chamber 330. Although examples of isolation chambers are described herein, it should be understood that the transfer device 305 and / or the housing 310 can have isolation chambers arranged in any suitable manner, and thus the isolation chamber 330 is not intended to be limited to the isolation chambers illustrated and described herein.

[0191] The isolation chamber 330 can have any suitable volume and / or fluid capacity. For example, in some embodiments, the isolation chamber 330 can have a volume and / or fluid capacity of from about 0.1 mL to about 5.0 mL. In some embodiments, the isolation portion 330 can have a volume measured with respect to the amount of body fluid (e.g., an initial or first amount of body fluid) configured to be transferred into the isolation chamber 330. For example, in some embodiments, the isolation chamber 330 can have a volume sufficient to receive a small initial body fluid of less than a microliter (e.g., a small volume such as 20 drops of body fluid, 10 drops of body fluid, 5 drops of body fluid, 1 drop of body fluid, or any suitable volume therebetween). In other embodiments, the isolation chamber 330 can have a volume sufficient to receive, for example, an initial volume of body fluid of about 5.0 mL, 10.0 mL, 15.0 mL, 20.0 mL, 30.0 mL, 40.0 mL, 50.0 mL, or more. In some embodiments, the isolation chamber 330 can have a volume equal to at least a portion of the volume of one or more lumens (e.g., the lumen of a needle, the inlet 312, and at least a portion of the fluid flow path 315) that place the isolation chamber 330 in fluid communication with the body fluid source. In still other embodiments, the isolation chamber 330 can have a volume based at least in part on the desired volume of body fluid to be used within or by the rapid diagnostic test device 370.

[0192] As shown in FIG. 4, the device 305 includes a flow controller 340 that is at least partially disposed within the housing 310 and configured to control, direct, and / or otherwise facilitate a selective flow of fluid through at least a portion of the housing 310, at least a portion of the fluid flow path 315, and / or at least a portion of the isolation chamber 330. In this regard, the fluid flow can be a liquid such as, for example, water, oil, a moistened fluid, a body fluid, and / or any other suitable liquid, and / or a gas such as air, oxygen, carbon dioxide, helium, nitrogen, ethylene oxide, and / or any other suitable gas.

[0193] The flow controller 340 can be of any suitable shape, size, and / or configuration. In some embodiments, the flow controller 340 can be, for example, a valve, a membrane, a diaphragm, a bladder, a plunger, a piston, a bag, a pouch, and / or any other suitable member having a desired rigidity, flexibility, and / or hardness, or any suitable combination thereof. In some embodiments, the flow controller 340 can be, for example, a restrictor, a vent, an absorption member, a selectively permeable member (e.g., a fluid-impermeable barrier or seal that at least selectively allows the passage of air or gas), a port, a junction, an actuator, etc., or any suitable combination thereof. In some embodiments, the flow controller 340 can be the same as or substantially the same as the flow controllers described in U.S. Patent No. 11,076,787, entitled "Fluid Control Devices and Methods of Using the Same," filed on September 12, 2018 (the " '787 Patent"), U.S. Patent Application Publication No. 2019 / 0365303, entitled "Fluid Control Devices and Methods of Using the Same," filed on May 30, 2019 (the " '303 Publication"), and / or U.S. Patent Application Publication No. 2020 / 0289039, entitled "Fluid Control Devices and Methods of Using the Same," filed on March 11, 2020 (the " '039 Publication"), the disclosures of each of these patents and patent applications being incorporated herein by reference in their entireties.

[0194] For example, in some embodiments, the transfer device 305 can be configured to selectively transfer a volume of body fluid to the isolation chamber 330 or to the outlet 313, based at least in part on a pressure differential between two or more portions of the transfer device 305. For example, the pressure differential can result from fluidly connecting the outlet 313 to the fluid collection device 395, which can define and / or be configured to generate a negative pressure (e.g., a vacuum reservoir, syringe, pressure filling cannister, and / or other source or potential energy for generating a vacuum or pressure differential). In other embodiments, the pressure differential can result from a change in volume and / or temperature. In yet other embodiments, the pressure differential can result from the transfer device 305, the housing 310, at least a portion of the actuator 350, and / or each part of the fluid flow path 315 being evacuated and / or filled (e.g., the isolation chamber 330 and / or any other suitable portion). In some embodiments, the pressure differential can be established automatically, or via direct or indirect intervention (e.g., by a user).

[0195] In some embodiments, the flow controller 340 can be configured to facilitate the movement of air (or other fluid) through one or more portions of the transfer device 305, which in some instances can enable or result in a pressure differential and / or pressure equalization across one or more portions of the housing 310. Further, the flow of fluid (e.g., gas and / or liquid) resulting from the pressure differential can be selectively controlled via the flow controller 340. For example, the flow controller 340 can be configured to transition between one or more operating states or conditions to control the flow of the fluid. In some embodiments, the flow controller 340 can be a member or device formed from an absorbent or semi-permeable material configured to selectively permit the flow of fluid therethrough. For example, such an absorbent material can transition from a first state, as described in detail in the '117 publication and / or the '303 publication, where the material permits the flow of gas (e.g., air) therethrough but blocks the flow of liquid (e.g., body fluid) therethrough, to a second state where the material substantially blocks the flow of gas and liquid therethrough (e.g., the flow controller 340 can be a selectively permeable blood barrier).

[0196] In some embodiments, the flow controller 340 can be configured to transition from a first state to a second state in response to a negative pressure differential and / or suction force exerted on at least a portion of the flow controller 340. For example, the flow controller 340 can include one or more valves, membranes, diaphragms, and the like. For example, the flow controller 340 can be in a first state (e.g., a storage or non-use state) prior to using the device 305, and in response to the outlet 313 being fluidly coupled to a fluid collection device 395 (e.g., a collection device configured to define or be configured to define a negative pressure and / or suction force), the flow controller 340 can be transitioned to the second state. In some embodiments, the flow controller 340 can be a bladder configured to transition or "flip" from a first state to a second state in response to a negative pressure differential and / or suction force exerted on the surface of the bladder as described in detail in Application Publication No. '303 and / or Application Publication No. '039.

[0197] In some embodiments, the size, shape, arrangement configuration, and / or constituent material of the flow controller 340 can be configured such that and / or selected in other ways such that the flow controller 340 transitions from the first state to the second state in a predetermined manner and / or at a predetermined or desired rate. In some examples, when controlling the rate at which the flow controller 340 transitions from the first state to the second state, the rate of body fluid flow into the isolation chamber 330 and / or the magnitude of the suction force generated within the isolation chamber 330 that acts to draw an initial volume of body fluid into the isolation chamber 330 can then be controlled and / or adjusted. Although not shown in FIG. 4, in some embodiments, the housing 310 and / or the flow controller 340 can include any suitable member, feature, opening, etc. configured to adjust the suction force applied onto or through the flow controller 340, whereby the rate at which the flow controller 340 transitions from the first state to the second state can be adjusted. In some examples, by controlling the rate at which the flow controller 340 transitions and / or the magnitude of the pressure difference and / or suction force generated within the isolation chamber 330, the hemolysis of a blood sample and / or the likelihood of vein collapse can be reduced, for example, which is particularly important when obtaining a body fluid sample from a vulnerable patient. In some examples, by adjusting the transition of the flow controller 340 and / or the pressure difference generated within the isolation chamber 330, the amount or volume of body fluid transferred into the isolation chamber 330 can be at least partially controlled (i.e., the volume of the initial amount of body fluid can be controlled).

[0198] In some embodiments, the flow controller 340 can include any suitable combination of devices, members, and / or features. It should be understood that the flow controllers included in the embodiments described herein are presented by way of example and not limitation. Thus, while a particular flow controller is described herein, it should be understood that fluid flow can be controlled through the transfer device 305 in any suitable manner.

[0199] The actuator 350 of device 305 is at least partially disposed within the housing 310 and is configured to control, direct, and / or otherwise facilitate a selective flow of fluid through at least a portion of the housing 310 and / or at least a portion of one or more fluid flow paths 315. The actuator 350 can be of any suitable shape, size, and / or configuration. In some embodiments, the actuator 350 can be a member or device configured to transition between any number of states in any suitable manner. Additionally, the actuator 350 can be actuated in any suitable manner (e.g., user-actuated, automatically actuated, mechanically actuated, electronically actuated, chemically actuated, etc.). For example, the actuator 350 can be similar to and / or substantially the same as any of the actuators described above with reference to actuator 250.

[0200] In the embodiment shown in FIG. 4, the actuator 350 can be configured to selectively establish fluid communication between the inlet 312 and the isolation chamber 330 when in the first state, and to selectively establish fluid communication between the inlet 312 and the outlet 313 when in the second state. When in the first state, the actuator 350 can be configured to allow body fluid to flow from the inlet 312 through at least a portion of the fluid flow path 315 into the isolation chamber 330 or to flow into the isolation chamber 330. In some embodiments, the actuator 350 can be configured to isolate, separate, segregate, and / or otherwise block fluid communication between the outlet 313 and the inlet 312, at least a portion of the fluid flow path 315, and / or the isolation chamber 330. When in the second state, the actuator 350 can be configured to allow subsequent volumes of body fluid (e.g., volumes of body fluid after an initial volume of body fluid) to be transferred from the inlet 312 through at least a portion of the fluid flow path 315 to the outlet 313 (and / or a fluid collection device 395 fluidly coupled to the outlet 313), as described in more detail herein. Additionally, when in the second state, the actuator 350 can be configured to isolate, separate, segregate, and / or otherwise block fluid communication between the isolation chamber 330 and the inlet 312, the outlet 313, and / or at least a portion of the fluid flow path 315. In the embodiment shown in FIG. 4, the transfer device 305 is a transfer device 305 such that the actuator 350 and the flow controller 340 collectively control the flow of fluid (e.g., gas and / or liquid) through the device, as described in more detail herein.

[0201] The rapid diagnostic test device 370 (also referred to herein as the "rapid test device") can be of any suitable shape, size, and / or configuration. In some embodiments, the rapid test device 370 can be removably coupled to the transfer device 305 or any suitable portion thereof. For example, in the embodiment shown in FIG. 4, the rapid test device 370 can be at least fluidly coupled to the isolation chamber of the transfer device 305. In other embodiments, the rapid test device 370 can be integrated into the transfer device 305 such that the rapid test device 370 is in fluid communication with the isolation chamber 330. For example, the transfer device 305 and the rapid test device 370 can be integrally or monolithically formed and / or integrated in other ways. In still other embodiments, the housing 310 can include ports, adapters, and / or receiving portions that can connect or insert the rapid test device 370 to establish fluid communication between the rapid test device 370 and the isolation chamber 330 and / or can be formed to do so.

[0202] In some such embodiments, coupling the rapid test device 370 to the transfer device 305 can act to transition one or more flow controllers, valves, partitions, ports, seals, etc. from a closed or sealed state to an open state to enable fluid communication between the transfer device 305 and the test device 370. Although not shown in FIG. 4, in some embodiments, the transfer device 305 can include a second actuator or the like that can be operated to establish fluid communication between the isolation chamber 330 and the rapid test device 370. In other embodiments, the actuator 350 can be transitioned to establish fluid communication between the isolation chamber 330 and the rapid test device 370.

[0203] In some implementations, the rapid test device 370 can be configured to receive a first quantity of body fluid from the transfer device 305 and perform one or more tests, assays, and / or diagnostic methods using the first quantity of body fluid. The rapid test device 370 can be any suitable test device. For example, the rapid test device 370 can be an LFA such as described in detail above with reference to the LFA170A shown in FIG. 2A or the LFA170B shown in FIG. 2B. In some implementations, the test device 370 and / or aspects or portions thereof can be substantially similar to the rapid test devices 170 and / or 270 described in detail above. Accordingly, the transfer device 370 and / or aspects or portions thereof are not described in further detail herein.

[0204] As described above, the system 300 can be used to obtain one or more volumes of body fluid from a patient that can be used in one or more tests, assays, and / or diagnostic methods. For example, in some instances, a user such as a physician, internist, nurse, phlebotomist, technician, etc. can operate the device 305 to establish fluid communication between the inlet 312 and a body fluid source (e.g., a patient's vein, cerebrospinal fluid (CSF) from the spinal cavity, urine collection, etc.). In some instances, the actuator 350 can be in a first state in which the inlet 312 is in fluid communication with a body fluid source (e.g., a part of a patient) such that at least a portion of the fluid flow path 315 establishes fluid communication between the inlet 312 and the isolation chamber 330.

[0205] Accordingly, the transfer device 305 can be configured to transfer an initial volume of body fluid from a body fluid source (e.g., a patient) to the rapid test device 370. More specifically, in the embodiment shown in FIG. 4, when the inlet 312 is placed in fluid communication with a body fluid source (e.g., a part of a patient), the outlet 313 can be fluidly connected to the fluid collection device 395. As described above, the fluid collection device 395 can be any suitable reservoir, container, and / or collection system configured to receive a volume of body fluid. For example, the fluid collection device 395 can be a vacuum reservoir or container that defines a negative pressure and / or can be a syringe that is operated to generate a negative pressure. In some examples, by connecting the outlet 313 to the fluid collection device 395, at least a portion of the fluid flow path 315 is selectively exposed to the negative pressure and / or suction force within the fluid collection device 395. In some implementations, the actuator 350 can be in a first state such that the outlet 313 is isolated from the inlet 312. Additionally, when the actuator 350 is in the first state, the outlet 313 can be in fluid communication with the flow controller 340 (e.g., via a portion of the fluid flow path 315). The flow controller 340 can similarly be in a first state of the flow controller 340 when the fluid collection device 395 is connected to the outlet 313.

[0206] In an embodiment where the flow controller 340 is a selectively permeable member or membrane, the arrangement of the flow controller 340 and the actuator 350 can be an arrangement that allows air or gas flow to pass through the flow controller 340 between the outlet 313 and the isolation chamber 330. In such an embodiment, this arrangement causes at least a portion of the negative pressure difference or suction force generated by the fluid collection device 395 to be transmitted into and / or through the isolation chamber 330, and then, as described in detail in the '117 application publication and / or the '303 application publication, act to draw an initial volume of body fluid from the body fluid source into the isolation chamber 330 through the inlet 312 and at least a portion of the fluid flow path 315.

[0207] Alternatively, in an embodiment where the flow controller 340 is a diaphragm, flap, valve, sleeve, etc., the arrangement of the flow controller 340 and the actuator 350 can be such that a part and / or surface of the flow controller 340 is in fluid communication with the outlet 313 (e.g., via a part of the fluid flow path 315). Thus, a negative pressure and / or suction force can be applied to the part and / or surface of the flow controller 340, and the flow controller 340 can act to transition the isolation chamber 330 from a first state where the isolation chamber 330 has a first volume to a second state where the isolation chamber 330 has a second volume greater than the first volume. The isolation chamber 330 can be an isolation chamber that acts to generate a negative pressure difference such that an increase in volume results in a decrease in pressure within the isolation chamber 330, thereby drawing body fluid into the isolation chamber 330. Thus, in such an embodiment, in response to the transition of the flow controller 340, an initial volume of body fluid can be drawn into the isolation chamber 330 (e.g., as a result of the volume of the isolation chamber 330 increasing as the flow controller 340 transitions from the first state to the second state), as described in detail in the '303 publication and / or the '039 publication.

[0208] The initial volume of the body fluid can be any suitable volume of body fluid, such as the volume or amount described above. For example, in some instances, the transfer device 305 can remain in a first state or configuration until a predetermined and / or desired volume (e.g., the initial volume) of body fluid is transferred to the isolation chamber 330. In some embodiments, the initial volume can be associated with and / or at least partially based on the volume of the isolation chamber 330 or a portion thereof (e.g., a volume sufficient to fill the isolation chamber 330 or a desired portion of the isolation chamber 330). In some embodiments, the initial volume can be associated with and / or at least partially based on a desired volume sufficient for the rapid test device 370 to perform one or more tests or assays. In other embodiments, the initial volume of body fluid can be associated with and / or at least partially based on an amount or volume of body fluid greater than the volume associated with the fluid flow path defined between the body fluid source and the isolation chamber 330. In yet other embodiments, the transfer device 305 can be configured to transfer a certain flow rate of body fluid (e.g., the initial volume) to the isolation portion 330 until the pressure differential between the isolation chamber 330 and the inlet 312 or the body fluid source is brought to a substantially equilibrium state and / or otherwise reduced below a desired threshold value.

[0209] In some embodiments, the transfer device 305 may be configured to transfer a volume of body fluid (e.g., an initial volume) into the isolation chamber 330 until the flow controller 340 transitions to its second configuration. In other words, in some embodiments, transferring an initial volume of body fluid into the isolation chamber 330 may act to place the flow controller 340 in its second state or configuration. For example, in embodiments where the flow controller 340 is a selectively permeable member, transferring an initial volume of body fluid into the isolation chamber 330 may be such that at least a portion of the initial volume wets and / or saturates the flow controller 340 and then causes the flow controller 340 to transition to its second state, as described in detail in the '117 application and / or the '303 application publication. In embodiments where the flow controller 340 is a diaphragm or the like, as described in detail in the '303 application publication and / or the '039 application publication, the transfer of the initial volume into the isolation chamber 330 may occur substantially simultaneously with the flow controller 340 being placed in its second state and / or configuration (e.g., in response to a negative pressure being generated by the fluid collection device 395). Further, in the embodiment shown in FIG. 4, the arrangement of the flow controller 340 is such that when the flow controller 340 is in its second state and / or configuration, it isolates and / or fluidly separates the isolation chamber 330 from the outlet 313 so that the negative pressure and / or suction force generated by the fluid collection device 395 no longer acts on the isolation chamber 330.

[0210] In some implementations, when the flow controller 340 is in its second state and before the actuator transitions from its first state to its second state, at least a portion of the initial volume of the body fluid can be transferred from the isolation chamber 330 into the rapid test device 370. In some embodiments, the actuator 350 is configured to isolate the isolation chamber 330 from at least a portion of the inlet 313, the outlet 315, and the fluid flow path 315. In such embodiments, a portion of the initial volume of the body fluid can be transferred from the isolation chamber 330 before, during, and / or after transitioning the actuator 350 from its first state to its second state. In some implementations, the transfer of a portion of the initial volume can be automatic. In other implementations, the transfer of a portion of the initial volume can be performed in response to one or more user inputs, etc.

[0211] In some embodiments, by transferring a portion of the initial volume of the body fluid to the rapid test device 370, testing and / or analysis of a portion of the initial volume of the body fluid can be initiated, as described in detail above with reference to the rapid test device 270. Further, the rapid test device 370 can be configured to perform any suitable test and / or assay. For example, the rapid test device 370 can be an LFA configured to test for the presence of lactate, IL6, PCT, CD64, and / or NNM, as described in detail above. Further, when the test or assay is complete, the rapid test device 370 can be configured to output the test result, which can be detected and / or determined by a human and / or one or more electronic devices, as described in detail above.

[0212] In some embodiments, the transition of the actuator 350 from its first state to its second state (e.g., placing the transfer device 305 in its second state or configuration) can isolate, separate, and / or retain an initial volume of body fluid within the isolation chamber 330 and / or the rapid test device 370. In other words, the actuator 350 can isolate and / or separate the isolation chamber 330 from one or more portions of the inlet 312, the outlet 313, and the fluid flow path 315. In some examples, isolating an initial volume of body fluid within the isolation chamber 330 can also isolate contaminants within the initial volume. Further, the arrangement of the rapid test devices 370 can be such that the tests and / or assays performed by the rapid test devices 370 are not affected by such contamination, which means that the accuracy of the test results output by the rapid test devices 370 is not affected by such contamination, as detailed above.

[0213] In addition to isolating the isolation chamber 330 from at least a portion of the inlet 312, the outlet 313, and the fluid flow path 315, placing the actuator 350 in its second state (and setting the flow rate controller 340 to its second state) also establishes fluid communication between the inlet 312 and the outlet 313 through at least a portion of the fluid flow path 315. For example, in some embodiments, transitioning the actuator 350 from its first state to its second state can, for example, open or close a port or valve, move one or more seals, move or remove one or more obstacles, define one or more portions of the flow path, and the like. Thus, in response to the negative pressure and / or suction force generated by the fluid collection device 395, one or more subsequent volumes of body fluid can flow from the inlet 312, through at least a portion of the fluid flow path 315, through the outlet 313, and into the fluid collection device 395. As described above, by isolating an initial volume of body fluid (e.g., within the isolation chamber 370) prior to collecting or obtaining one or more subsequent volumes of body fluid, the amount of contaminants in the one or more sample volumes is reduced and / or substantially eliminated. Thus, the system 300 can be configured to obtain an initial volume of body fluid for use in rapid tests that are relatively insensitive to contamination, and subsequent volumes of body fluid for use in tests that are relatively sensitive to contamination, as described above with reference to systems 100 and / or 200.

[0214] Figures 5A and 5B are schematic diagrams of a fluid transfer and assay system 400 according to an embodiment, shown in a first state and a second state, respectively. The fluid transfer and assay system 400 (also referred to herein as the "system") can include at least a fluid transfer device 405 and a rapid diagnostic test device 470. Some parts and / or aspects of the fluid transfer device 405 and / or the rapid diagnostic test device 470 can be the same as and / or substantially the same as the fluid transfer devices 105, 205, and / or 305, and / or the rapid diagnostic test devices 170 (and / or LFA 170A, 170B), 270, and / or 370 described in detail above, respectively. Therefore, some such parts and / or aspects will not be described in further detail herein.

[0215] The fluid transfer device 405 (also referred to herein as the "transfer device") can be of any suitable shape, size, and / or configuration. In some implementations, the transfer device 405 can be configured to draw a body fluid (e.g., blood) from a patient into the transfer device 405 and / or through the transfer device 405. Additionally, the transfer device 405 can be configured to transfer at least a portion of the drawn body fluid to one or more other devices, reservoirs, containers, vials, machines, tests, assays, etc., such as the rapid diagnostic test device 470 and / or one or more fluid collection devices (not shown in Figures 5A and 5B).

[0216] The transfer device 405 includes at least a housing 410 and an actuator 450. The housing 410 of the device 405 can be of any suitable shape, size, and / or configuration. For example, in some embodiments, the housing 410 can be similar to and / or substantially the same as the housings 210 and / or 310 described above. Specifically, the housing 410 has and / or forms an inlet 412 and an outlet 413, and can define at least one fluid flow path (not shown in FIGS. 5A and 5B) between the inlet 412 and the outlet 413. The inlet 412 can be any suitable inlet or port and can be configured to establish fluid communication between the housing 410 and a body fluid source (e.g., a patient). The outlet 413 can be any suitable outlet or port and can be configured to establish fluid communication between the housing 410 and a fluid collection device (not shown in FIGS. 5A and 5B), such as any of the fluid collection devices described in detail above. The one or more fluid flow paths defined by the housing 410 extend between the inlet 412 and the outlet 413 and can selectively establish fluid communication between the inlet 412 and the outlet 413, as described in more detail herein.

[0217] As described above with reference to housing 310, housing 410 shown in FIGS. 5A and 5B includes, forms, and / or is coupled to an isolation chamber 430 configured to be placed in selective fluid communication with a fluid flow path and / or at least an inlet 412. Additionally, isolation chamber 430 includes, is coupled to, and / or is otherwise configured to be in fluid communication with a rapid diagnostic test device 470. Isolation chamber 430 can have any suitable shape, size, and / or configuration. For example, in some embodiments, isolation chamber 430 can have a volume and / or fluid capacity of from about 0.1 mL to about 5.0 mL. In some embodiments, isolation chamber 430 can have a volume measured with respect to an amount of body fluid (e.g., an initial or first amount of body fluid) configured to be transferred into isolation chamber 430 and / or tested by rapid diagnostic test device 470. In some embodiments, isolation chamber 430 and / or at least a portion thereof can be substantially similar to isolation chamber 330 described above with reference to FIG. 4, at least in form and / or function. Thus, some portions and / or aspects of isolation chamber 430 are not described in further detail herein.

[0218] In the embodiments shown in FIGS. 5A and 5B, at least a portion of the isolation chamber 430 can include an absorbent and / or hydrophilic material 431. Additionally, the isolation chamber 430 includes a sampling portion 435 and a vent hole 424. The absorbent material 431 can be disposed within a portion of the isolation chamber 430. For example, one or more inner surfaces of the isolation chamber 431 can be lined with and / or formed by the absorbent material 431. As shown in FIGS. 5A and 5B, the arrangement of the isolation chamber 430 can be such that the sampling portion 435 of the isolation chamber 430 is downstream of the absorbent material 431 (e.g., downstream with respect to a portion of the isolation chamber 430 being temporarily fluidly connected to the inlet 412). In this way, the absorbent material 431 can be configured to receive and / or absorb a first portion or part of the initial volume of body fluid transferred into the isolation chamber 430. In some implementations, the absorbent material 431 can become saturated after absorbing a predetermined amount or volume of body fluid, whereby any additional amount or volume of body fluid transferred into the isolation chamber 430 can flow into the sampling portion 435. As described in more detail herein, the sampling portion 435 of the isolation chamber 430 can be placed in fluid communication with the rapid diagnostic test device 470 to transfer a portion of the initial volume of body fluid disposed within the sampling portion 435 into the rapid diagnostic test device 470.

[0219] Vent hole 424 is connected to housing 410 and / or isolation chamber 430 and is in fluid communication with the internal volume of isolation chamber 430. Vent hole 424 can be configured to release an air or gas flow from isolation chamber 430 and / or otherwise enable it when an initial volume of body fluid is transferred into isolation chamber 430. In some implementations, releasing air or gas from isolation chamber 430 (e.g., via vent hole 424) can reduce the amount of pressure within isolation chamber 430 and, in other cases, can limit and / or impede the flow of body fluid into isolation chamber 430. In some implementations, releasing air or gas through vent hole 424 can enable a negative pressure differential that can facilitate the transfer of an initial volume of body fluid into isolation chamber 430. Although absorbent material 431 and vent hole 424 are shown as separate components in FIGS. 5A and 5B, in other embodiments, absorbent material 431 can be configured to vent isolation chamber 430 and can form one or more vent holes configured to absorb an initial volume of a first portion or part thereof. For example, absorbent material 431 can form one or more walls of isolation chamber 430 or one or more portions of the walls.

[0220] The actuator 450 of device 405 can be of any suitable shape, size, and / or configuration. In some embodiments, the actuator 450 and / or aspects or portions thereof can be similar to and / or substantially the same as the actuators 150, 250, and / or 350 described in detail above. In some embodiments, the actuator 450 can be at least partially disposed within and / or at least partially formed by the housing 410. As described above, the actuator 450 can be configured to control, direct, and / or otherwise facilitate a selective flow of fluid through at least a portion of the housing 410 and / or at least a portion of one or more fluid flow paths. In some embodiments, the actuator 450 can be a member or device configured to transition between any number of states (e.g., two, three, four, or more) in any suitable manner (e.g., user-actuated, auto-actuated, mechanically actuated, electronically actuated, chemically actuated, etc.).

[0221] More specifically, in the embodiments shown in FIGS. 5A and 5B, the actuator 450 can be configured to transition between a first state (FIG. 5A) in which the inlet 412 is in fluid communication with the isolation chamber 430 and a second state (FIG. 5B) in which the inlet 412 is in fluid communication with the outlet 413. In some embodiments, the actuator 450 can be configured to isolate, separate, insulate, and / or otherwise prevent fluid communication between the outlet 413 and the inlet 412 and / or between the outlet 413 and the isolation chamber 430 when in the first state. Conversely, in the second state, the actuator 450 can be configured to allow a subsequent volume of body fluid (e.g., a volume of body fluid after an initial volume of body fluid) to be transferred from the inlet 412 through one or more fluid flow paths (not shown in FIGS. 5A and 5B) to the outlet 413 (and / or a fluid collection device fluidly coupled to the outlet 413). Additionally, when in the second state, the actuator 450 can be configured to isolate, separate, insulate, and / or otherwise prevent fluid communication between the isolation chamber 430 and the inlet 412, between the isolation chamber 430 and the outlet 413, and / or between the isolation chamber 430 and at least a portion of the fluid flow path extending between the inlet 412 and the outlet 413. Accordingly, the actuator 450 can be structurally and / or functionally similar to the actuators 150, 250, and / or 350 described in detail above.

[0222] The rapid diagnostic test device 470 (also referred to herein as the "rapid test device" or simply the "test device") can be of any suitable shape, size, and / or configuration. In some embodiments, the rapid test device 470 can be removably coupled to the transfer device 405 or any suitable portion thereof. For example, in the embodiments shown in FIGS. 5A and 5B, the rapid test device 470 can be configured to engage or couple with the housing 410 and / or the isolation chamber 4350 such that the rapid test device 470 is placed in fluid communication with the sampling portion 435 of the isolation chamber 430. In some embodiments, the housing 410 can include, and / or can form, ports, adapters, and / or receiving portions to which the rapid test device 470 can be coupled or into which the rapid test device 470 can be inserted to establish fluid communication between the rapid test device 470 and the sampling portion 435 of the isolation chamber 430. Further, in the embodiments shown in FIGS. 5A and 5B, fluid communication can be established between the isolation chamber 430 and the rapid test device 470 by transitioning the actuator 450 from its first state to its second state (e.g., fluid communication can be established via one or more flow controllers, valves, diaphragms, ports, seals, aligned flow paths, and / or other suitable members or devices for establishing fluid communication). In some embodiments, fluid communication can be established between the isolation chamber 430 and the rapid test device 470 by transitioning the actuator 450 from its first state to its second state.

[0223] In some implementations, the rapid test device 470 can be configured to receive a first quantity of body fluid from the sampling portion 435 of the isolation chamber 430 and perform one or more tests, assays, and / or diagnostic methods using the first quantity of body fluid. The rapid test device 470 can be any suitable test device. For example, the rapid test device 470 can be an LFA such as those described in detail above with reference to the LFA170A shown in FIG. 2A or the LFA170B shown in FIG. 2B. The dashed lines in FIGS. 5A and 5B indicate the possibility of including at least one additional capture element when two or more biomarkers are measured. In some implementations, the test device 470 and / or aspects or portions thereof can be substantially similar to the rapid test devices 170, 270, and / or 370 described in detail above. Accordingly, the transfer device 470 and / or aspects or portions thereof are not described in further detail herein.

[0224] System 400 can be used to obtain one or more volumes of body fluid from a patient for use in one or more tests, assays, and / or diagnostic methods. As described above, for example, inlet 412 can be placed in fluid communication with a body fluid source. In some examples, actuator 450 can be in a first state when inlet 412 is placed in fluid communication with a body fluid source (e.g., a part of a patient), as shown in FIG. 5A, thereby establishing fluid communication between inlet 412 and isolation chamber 430 and isolating outlet 413 from inlet 412. Thus, transfer device 405 can be configured to transfer an initial volume of body fluid from a body fluid source (e.g., a patient) to rapid test device 470. In some implementations, the initial volume of body fluid can flow into and / or enter isolation chamber 430 in response to a pressure differential between isolation chamber 430 and inlet 412 and / or the body fluid source. In some embodiments, vent 424 can be configured to allow air or gas to flow from isolation chamber 430 to the exterior, thereby facilitating the flow of the initial volume of body fluid into isolation chamber 430. In some embodiments, vent 424 can be configured to vent isolation chamber 430 in a manner similar to the vents and / or the like described in, for example, the '117 publication.

[0225] The initial volume of body fluid can be any suitable volume of body fluid, such as the volume or amount described above. More specifically, in the embodiments shown in FIGS. 5A and 5B, the initial volume of body fluid saturates and / or wets (or substantially saturates and / or wets) the absorbent material 431 disposed within the isolation chamber 430 and fills (or substantially fills) the sampling portion 435 of the isolation chamber 430. In some embodiments, the filling of the isolation chamber 430 can be continuous in that the flow of the initial volume of body fluid is first absorbed by the absorbent material 431 until the absorbent material 431 is saturated, and then the remaining portion of the initial volume of body fluid can flow into and / or fill the sampling portion 435 of the isolation chamber 430. In some implementations, the continuous filling of the isolation chamber 430 can be such that a portion (e.g., a first portion) of the initial volume of body fluid can contain contaminants (e.g., contaminants associated with and / or resulting from a venipuncture event, fluid connections of one or more components, etc.), while a portion (e.g., a second portion) of the initial volume of body fluid can contain a reduced amount of contaminants and / or can be substantially free of contaminants. In some examples, when the initial volume of body fluid is transferred into the isolation chamber 430, the flow of the body fluid can be stopped and / or the pressure differential can be substantially equalized, thereby decelerating or stopping the flow of the body fluid.

[0226] In some embodiments, after transferring an initial volume of body fluid into the isolation chamber 430, the actuator 450 can transition from its first state (FIG. 5A) to its second state (FIG. 5B). For example, in some embodiments, the actuator 450 can be moved, slid, switched, rotated, and / or otherwise transitioned relative to the inlet 412 and the outlet 413. In some embodiments, transitioning and / or moving the actuator 450 can include transitioning and / or moving at least a portion of the housing 410. In other embodiments, the actuator 450 can be moved relative to the housing 410 (e.g., there is no need to transition and / or move the housing 410).

[0227] As shown in FIG. 5B, by transitioning the actuator 450 from the first state to the second state, fluid communication can be established between the sampling portion 435 of the isolation chamber 430 and the rapid test device 470, and the isolation chamber 430 can be isolated from the inlet 412, the outlet 413, and / or one or more portions of the fluid flow path therebetween. In some embodiments, the arrangement of the actuator 450 is such that placing the actuator 450 in the second state results in and / or increases a void between a portion of the isolation chamber 430 that includes the absorbent material 431 and a portion of the isolation chamber 430 that includes, forms, and / or defines the sampling portion 435. The void can facilitate the transfer of body fluid from the sampling portion 435 to the rapid test device 470 (e.g., by enabling a desired relative pressure or pressure differential). Additionally, in examples where contaminants are included in a portion of the initial volume absorbed by the absorbent material 431, such an arrangement can ensure that only a portion of the initial volume disposed in the sampling portion 435 of the isolation chamber 430 is transferred to the rapid test device 470.

[0228] At least a portion of the initial volume of body fluid can be transferred into the rapid test device 470 from the sampling portion 435 of the isolation chamber 430 when the actuator 450 transitions from its first state to its second state. In some implementations, the transfer of a portion of the initial volume can be automatic. In other implementations, the transfer of a portion of the initial volume can be performed in response to one or more user inputs, for example. In some implementations, by placing the actuator 450 in the second state, the rapid test device 470 can be fluidly coupled to the sampling portion 435 of the isolation chamber 430, thereby enabling fluid transfer between the rapid test device 470 and the sampling portion 435.

[0229] In some embodiments, by transferring a portion of the initial volume of body fluid to the rapid test device 470, an examination and / or assay of a portion of the initial volume of body fluid can be initiated as described in detail above with reference to the rapid test device 270. In some examples, the system 400, the transfer device 405, and / or the rapid test device 470 can be configured to provide a buffer 481 (or any other suitable solution) that can be mixed with a portion of the initial volume of body fluid, as shown in FIG. 5B. The rapid test device 470 can be configured to perform any suitable examination and / or assay. For example, the rapid test device 470 can be an LFA configured to examine for the presence of lactate, IL6, PCT, CD64, and / or NNM, as described in detail above. Further, when the examination or assay is complete, the rapid test device 470 can be configured to output test results that can be detected and / or determined by a human and / or one or more electronic devices, as described in detail above with reference to the rapid test devices 170, 270, and / or 370.

[0230] By transitioning the actuator 450 from its first state to its second state, an initial volume of body fluid can be isolated, separated, and / or retained within the isolation chamber 430 and / or the rapid test device 470. In other words, the actuator 450 can isolate and / or separate the isolation chamber 430 from the inlet 412, the outlet 413, and one or more portions of the fluid flow path. In some examples, isolating an initial volume of body fluid within the isolation chamber 430 can also isolate contaminants in the initial volume (e.g., at least a portion of the initial volume absorbed by the absorbent material 431). Further, the arrangement of the rapid test device 470 can be such that the tests and / or assays performed by the rapid test device 470 are not affected by such contamination, which means that the accuracy of the test results output by the rapid test device 470 is not affected by such contamination, as detailed above. In other examples, by receiving and / or absorbing a first portion or part of the initial volume of body fluid by the absorbent material 431, the rapid test device 470 can be enabled to perform one or more tests that can be at least partially sensitive to contaminants.

[0231] In addition, transitioning the actuator 450 to its second state establishes fluid communication between the inlet 412 and the outlet 413 via at least a portion of the fluid flow path disposed between the inlet 412 and the outlet 413. For example, transitioning the actuator 450 from its first state to its second state can open or close a port or valve, move one or more seals, move or remove one or more obstacles, define one or more portions of the flow path, and the like. In some implementations, the outlet 413 can be placed in fluid communication with the fluid collection device before or after the actuator is placed in its second state. As described in detail above, the fluid collection device can define a negative pressure and / or suction force that acts to draw body fluid into the fluid collection device and / or can be configured to generate such negative pressure and / or suction force. Thus, in response to the negative pressure and / or suction force, one or more subsequent volumes of body fluid can flow from the inlet 412, through any suitable fluid flow path or portion thereof, through the outlet 413, and into the fluid collection device. Thus, as described above, isolating an initial volume of body fluid within the isolation chamber 430 prior to collecting or obtaining one or more subsequent volumes of body fluid reduces and / or substantially eliminates the amount of contaminants in the one or more subsequent volumes. Thus, the system 400 can acquire an initial volume of body fluid for use in one or more rapid assay processes and a subsequent volume of body fluid for use in an assay (e.g., a blood culture assay) that is relatively sensitive to contamination, as described above with reference to systems 100, 200, and / or 300.

[0232] Figures 6A - 6D are schematic diagrams of at least a part of a fluid transfer and assay system 500 according to one embodiment. The fluid transfer and assay system 500 (also referred to herein as the "system") can include at least a fluid transfer device 505 and a rapid diagnostic test device 570. Some parts and / or aspects of the fluid transfer device 505 and / or the rapid diagnostic test device 570 can be the same as and / or substantially the same as the fluid transfer devices 105, 205, 305, and / or 405, and / or the rapid diagnostic test devices 170 (and / or LFA 170A, 170B), 270, 370, and / or 470 described in detail above respectively. The dashed lines in Figures 6A - 6D indicate the possibility of including at least one additional capture element when two or more biomarkers are measured. Therefore, some such parts and / or aspects will not be described in further detail herein.

[0233] The fluid transfer device 505 (also referred to herein as the "transfer device") can be of any suitable shape, size, and / or configuration. In some implementations, the transfer device 505 can be configured to draw a body fluid (e.g., blood) from a patient into the transfer device 505 and / or through the transfer device 505. Additionally, the transfer device 505 can be configured to transfer at least a part of the drawn body fluid to one or more other devices, reservoirs, containers, vials, machines, tests, assays, etc., such as the rapid diagnostic test device 570 and / or one or more fluid collection devices (not shown in Figures 6A - 6D). The transfer device 505 and / or its aspects or parts can be substantially the same as any of the transfer devices 105, 205, 305, and / or 405 described in detail above. Therefore, the transfer device 505 will not be described in further detail herein.

[0234] The rapid diagnostic test device 570 (also referred to herein as the "rapid test device" or simply the "test device") can be any suitable test device. In some implementations, the test device 570 and / or aspects or portions thereof can be substantially similar to the rapid test devices 170 (and / or LFA 170A, 170B), 270, 370, and / or 470 described in detail above. Accordingly, the transfer device 570 and / or aspects or portions thereof will not be described in further detail herein.

[0235] In the embodiments shown in FIGS. 6A-6D, the rapid test device 570 can be, for example, an LFA as described in detail above with reference to the LFA 170A shown in FIG. 2A or the LFA 170B shown in FIG. 2B. The rapid test device 570 includes a substrate 571 having any suitable configuration, such as a capillary bed, as described in detail above. Additionally, the rapid test device 570 includes a connecting member 578 that can be coupled to the substrate 571 via an attachment mechanism 579. The connecting member 578 can be any suitable connecting member configured to establish fluid communication with the internal volume of the transfer device 505 in response to the rapid test device 570 being coupled to the transfer device 505. For example, as shown in FIGS. 6A and 6B, the rapid test device 570 and / or its connecting member 578 can be configured to couple to the transfer device 505 via a port 525 (e.g., any suitable port, vent, coupler, opening, valve, junction, etc.). In some embodiments, the connecting member 578 can be, for example, a piercing member, needle, tube, etc. that can pierce the port 525 and / or advance through the port 525 in other ways. In some embodiments, the connecting member 578 can be a capillary member configured to transfer fluid via capillary action. In some embodiments, the port 525 can have a self-sealing action that allows the port 525 to be sealed when the connecting portion 578 of the rapid test device 570 is removed from the port 525. In some embodiments, the port 525 and / or at least a portion thereof can include and / or form a vent similar to the vent 424.

[0236] The attachment mechanism 579 can be any suitable member, mechanism, device, etc. configured to attach the connecting member 578 to the substrate 571. In some embodiments, the attachment mechanism 579 can be configured to transition between two or more states or configurations to selectively place the connecting member 578 in fluid communication with a portion of the substrate 571 (e.g., a sample portion, element, and / or capillary bed). More specifically, the attachment mechanism 579 can be configured to transition between a first state and / or configuration (Figs. 6A-6C) and a second state and / or configuration (Fig. 6D).

[0237] When in the first state, the rapid test device 570 can be coupled to the transfer device 505, and the connection portion 578 can establish fluid communication with the internal volume of the transfer device 505 (e.g., via port 525). As shown in Fig. 6B, the connecting member 578 can receive at least a portion of the volume of body fluid disposed within the transfer device 505 (e.g., via capillary action, pressure differential, and / or any other fluid transfer mode). As shown in Figs. 6C and 6D, when the connecting member 578 has received the desired volume of body fluid, the rapid test device 570 can be disconnected from the transfer device 505, and the attachment mechanism 579 can be transitioned from its first state to its second state.

[0238] For example, in some embodiments, the attachment mechanism 579 can be a living hinge or the like that can be bent, folded, deformed, and / or reconfigured in other ways. When the attachment mechanism 579 is in the second state, the connecting member 578 can be in fluid communication with a portion of the substrate 571 (e.g., a sample portion, element, and / or capillary bed), as shown in FIG. 6D. Thus, the volume of body fluid contained in the connecting member 578 can be transferred to a portion of the substrate 571. Additionally, in some implementations, when the attachment mechanism 579 is in the second state, a buffer 581 and / or any other suitable solution can be transferred to the substrate 571. The buffer 581 can be transferred to the substrate 571 via the connecting member 578, any suitable portion of the attachment mechanism 579, and / or any other suitable portion of the rapid test device 570. Thus, the buffer 581 can be mixed with the volume of body fluid described above, and the mixture can flow along the substrate 571 for testing, as described in detail above. In some implementations, the rapid test device 570 can be configured to test for the presence of lactate, IL6, PCT, CD64, and / or NNM, which can indicate a patient's condition such as sepsis. Further, when the test or assay is complete, the rapid test device 570 can be configured to output test results that can be detected and / or determined by a human and / or one or more electronic devices, as described in detail above with reference to the rapid test devices 170, 270, 370, and / or 470.

[0239] Figures 7A - 7D are schematic diagrams of a fluid transfer and assay system 600 according to one embodiment. The fluid transfer and assay system 600 (also referred to herein as the "system") can include at least a fluid transfer device 605 and a rapid diagnostic test device 670. Some parts and / or aspects of the fluid transfer device 605 and / or the rapid diagnostic test device 670 can be similar to and / or substantially the same as the fluid transfer devices 105, 205, 305, 405, and / or 505, and / or the rapid diagnostic test devices 170 (and / or LFA 170A, 170B), 270, 370, 470, and / or 570 described in detail above. The dashed lines in Figure 7D indicate the possibility of including at least one additional capture element when two or more biomarkers are measured. Therefore, some such parts and / or aspects will not be described in further detail herein.

[0240] The fluid transfer device 605 (also referred to herein as the "transfer device") can be of any suitable shape, size, and / or configuration. In some implementations, the transfer device 605 can be configured to draw bodily fluid (e.g., blood) from a patient into the transfer device 605 and / or through the transfer device 605. Additionally, the transfer device 605 can be configured to transfer at least a portion of the drawn bodily fluid to one or more other devices, reservoirs, containers, vials, machines, tests, assays, etc., such as the rapid diagnostic test device 670 and / or one or more fluid collection devices (not shown in Figures 7A - 7D).

[0241] The transfer device 605 includes at least a housing 610 and an actuator 650. The housing 610 of the device 605 can be of any suitable shape, size, and / or configuration. For example, in some embodiments, the housing 610 can be similar to and / or substantially the same as the housings 210, 310, and / or 410 described above. Specifically, the housing 610 has and / or forms an inlet 612 and an outlet 613, and can define a fluid flow path 615 between the inlet 612 and the outlet 613. The inlet 612 can be any suitable inlet or port and can be configured to establish fluid communication between the housing 610 and a body fluid source (e.g., a patient). The outlet 613 can be any suitable outlet or port and can be configured to establish fluid communication between the housing 610 and a fluid collection device (not shown in FIGS. 7A-7D), such as any of the fluid collection devices described in detail above. The fluid flow path 615 defined by the housing 610 extends between the inlet 612 and the outlet 613 and can selectively establish fluid communication between the inlet 612 and the outlet 613, as described in more detail herein.

[0242] As described above with reference to the housing 410, the housing 610 shown in FIGS. 7A-7D includes, forms, and / or is connected to an isolation chamber 630 configured to selectively fluid communicate with a fluid flow path and / or at least the inlet 612. In addition, the isolation chamber 630 includes, forms, and / or defines a sampling portion 635 and a port 625. The isolation chamber 630 can have any suitable shape, size, and / or configuration. For example, in some embodiments, the isolation chamber 630 and / or at least a portion thereof can be substantially similar to the isolation chambers 330 and / or 430 described in detail above, at least in form and / or function. Therefore, some parts and / or aspects of the isolation chamber 630 are not described in more detail herein.

[0243] Port 625 is connected to housing 610 and / or isolation chamber 630 and is in fluid communication with the internal volume of isolation chamber 630. More specifically, as shown in FIGS. 7A-7D, port 625 is included in and / or connected to housing 610 and is in fluid communication with sampling portion 635 of isolation chamber 630. In some embodiments, port 625 and / or at least a portion thereof may be configured to release an air or gas flow from isolation chamber 630 and / or otherwise enable, as described in detail above with reference to vent hole 424, when an initial volume of body fluid is transferred into isolation chamber 630. Sampling portion 635 of isolation chamber 630 may be placed in fluid communication with rapid diagnostic test device 670 to transfer a portion of the initial volume of body fluid disposed within sampling portion 635 into rapid diagnostic test device 670. In the embodiments shown in FIGS. 7A-7D, for example, rapid diagnostic test device 670 may be placed in fluid communication with sampling portion 635 via port 625 and / or any other suitable port, as described above with reference to port 525 shown in FIGS. 6A and 6B.

[0244] Actuator 650 of device 605 can be of any suitable shape, size, and / or configuration. In some embodiments, actuator 650 and / or aspects or portions thereof may be similar to and / or substantially the same as actuators 150, 250, 350, and / or 450 described in detail above. In some embodiments, actuator 650 may be at least partially disposed within housing 610 and / or may be at least partially formed by housing 610. As described above, actuator 650 may be configured to control, direct, and / or otherwise facilitate a selective flow of fluid through at least a portion of housing 610 and / or at least a portion of one or more fluid flow paths. Actuator 650 can be any suitable member or device configured to transition between any number of states (e.g., two, three, four, or more) in any suitable manner (e.g., user-actuated, automated, mechanical, electrical, chemical, etc.).

[0245] More specifically, in the embodiments shown in FIGS. 7A-7D, the actuator 650 includes a first member 651 and a second member 660. The first member 651 of the actuator 650 can be of any suitable shape, size, and / or configuration. The first member 651 can be, for example, a plunger having at least one seal 652 (e.g., disposed at an end of the first member 651). In some embodiments, the end of the first member 651 can separate and / or at least partially define, for example, the sampling portion 635 of the isolation chamber 630. For example, the sampling portion 635 of the isolation chamber 630 can be disposed on one side of the end of the first member 651, and the remaining portion of the isolation chamber 630 can be disposed on the opposite side of the end of the first member 651. Further, the seal arrangement of the seal 652 can be such that the seal 652 engages and / or contacts the inner surface of the housing 610 to form and / or define a substantial fluid seal between the seal 652 and the housing 610. The first member 651 also includes one or more valves, ports, openings, channels, selective permeability members, etc. (referred to herein as "valve 653") configured to establish selective fluid communication between the sampling portion 635 of the isolation chamber 635 and the remaining portion of the isolation chamber 630, as will be described in more detail herein.

[0246] The second member 660 of the actuator 650 can be of any suitable shape, size, and / or configuration. For example, in the embodiments shown in FIGS. 7A-7D, the second member 660 can be disposed around and / or on at least a portion of the first member 651. The second member 660 includes a set of seals 661. More specifically, the second member 660 can include a set of three seals. As shown, the second member 660 can have a first end and a second end opposite the first end. The first end of the second member 660 includes an outer seal 661 configured to engage and / or contact the inner surface of the housing 610 to define a substantially fluid-tight seal therebetween. In addition, the first end of the second member 660 includes an inner seal 661 configured to engage and / or contact a portion of the first member 651 to define a substantially fluid-tight seal between the second member 660 and the first member 651. The second end of the second member 660 includes an outer seal 661 configured to engage and / or contact the inner surface of the housing to define a substantial fluid-tight seal between the second member 660 and the housing.

[0247] As shown in FIGS. 7A-7D, the arrangement of the first member 651 and the second member 660 of the actuator is such that a portion of the isolation chamber 630 (e.g., a portion other than the sampling portion 635) is disposed and / or defined, for example, between the end of the first member 651 and the first end of the second member 660. In addition, the second member 660 is configured to at least partially define a fluid flow path 615 between the first end and the second end of the second member 660. Thus, the first end of the second member 660 and the seals 661 included in the first end isolate and / or fluidly separate the isolation chamber 630 from the fluid flow path 615.

[0248] The actuator 650 is configured to transition between at least a first state, a second state, and a third state. As shown in FIGS. 7A-7D, the end of the first member 651 and the seal 652 included therein are disposed and maintained on the first side of the inlet 612 and the first side of the outlet 613 regardless of the state of the actuator 650. Similarly, the second end of the second member 660 and the seal member 661 included therein are disposed and maintained on the second side of the inlet 612 (opposite the first side) and the second side of the outlet 613 (opposite the first side) regardless of the state of the actuator 650. However, the first end of the second member 660 and the seal member 661 disposed thereon are (i) disposed on the second side of the inlet 612 and the first side of the outlet 613 when the actuator 650 is in the first and second states (FIGS. 7A and 7B), and (ii) disposed on the first side of the inlet 612 and the first side of the outlet 613 when the actuator 650 is in the third state (FIGS. 7C and 7D). Thus, the arrangement of the actuator 650 is such that, as described in more detail herein, the transition of the actuator 650 can selectively direct and / or divert the flow of fluid between (i) the inlet 612 and the isolation chamber 630 and (ii) the inlet 612 and the outlet 613 via the fluid flow path 615.

[0249] The rapid diagnostic test device 670 (also referred to herein as the "rapid test device" or simply the "test device") can be any suitable test device. For example, the rapid test device 670 can be an LFA such as described in detail above with reference to the LFA170A shown in FIG. 2A or the LFA170B shown in FIG. 2B. In some implementations, the test device 670 and / or aspects or portions thereof can be substantially similar to the rapid test devices 170, 270, 370, and / or 470 described in detail above. Accordingly, the transfer device 670 and / or aspects or portions thereof will not be described in further detail herein.

[0250] As shown in FIG. 7D, the rapid test device 670 can be configured to engage or connect with the housing 610 via the port 625. In some embodiments, for example, the port 625 is configured to (i) release air or gas from the isolation chamber 630 as described above with reference to the vent hole 424, and (ii) receive a portion of the rapid test device 670 and place the rapid test device 670 to be in fluid communication with the sampling portion 635 of the isolation chamber 630. It can be a valve, a coupler, and / or any suitable reconfigurable member or device. For example, the rapid test device 670 can include a connecting member 678 that can establish fluid communication with the sampling portion 635 of the isolation chamber 630 when the rapid test device 670 is connected to the sampling portion 635 of the isolation chamber 630. In some embodiments, the connecting member 678 can be, for example, a piercing member, a needle, a tube, a capillary, etc. that can pierce the port 625 and / or otherwise advance through the port 625. In some embodiments, the connecting member 678 can be substantially similar to the connecting member 578 described above with reference to FIGS. 6A-6D. In some embodiments, the port 625 can have a self-healing function that allows the port 625 to be sealed when the connecting portion 678 of the test device 670 is removed from the port 625. As shown in FIG. 7D, the connecting portion 678 of the test device 670 can be connected to the substrate 671 of the test device 670 (e.g., directly connected to the substrate 671 and / or connected via an attachment mechanism such as the attachment mechanism 579). In this way, the connecting portion 678 can transfer a volume of body fluid from the sampling portion 635 of the isolation chamber 630 into the test device 670. Accordingly, the test device 670 can perform one or more tests, assays, and / or diagnostic methods using the volume of the body fluid described above.

[0251] System 600 can be used to obtain one or more volumes of body fluid from a patient for use in one or more tests, assays, and / or diagnostic methods. As described above, for example, inlet 612 can be placed in fluid communication with a body fluid source. Actuator 650 can be in a first state when inlet 612 is placed in fluid communication with a body fluid source (e.g., a part of a patient), thereby establishing fluid communication between inlet 612 and isolation chamber 630 and isolating outlet 613 from inlet 612. Further, when actuator 650 is in the first state, the end of first member 651 can be near or adjacent to the first side of inlet 612, and the first end of second member 660 can be near or adjacent to the second side of inlet 612. In this way, the portion of isolation chamber 630 defined between first member 651 and second member 660 can have a first volume.

[0252] In some examples, when inlet 612 is placed in fluid communication with a body fluid source, actuator 650 can transition from its first state to its second state. For example, as shown in FIG. 7B, first member 651 can be transitioned or moved relative to inlet 612 and second member 660, thereby increasing the volume of the portion of isolation chamber 630 disposed between first member 651 and second member 660. Additionally, the transition and / or movement of first member 651 can decrease the volume of sampling portion 635 of isolation chamber 630, and the arrangement of ports 625 can be such that air or gas contained in sampling portion 635 can leak and / or flow out of sampling portion 635. The end of first member 651 can be configured to limit and / or substantially prevent the flow of air from sampling portion 635 of isolation chamber 630 to the remainder of isolation chamber 630, whereby the increase in volume within the remainder of isolation chamber 630 results in a negative pressure differential that acts when drawing an initial volume of body fluid from the body fluid source through inlet 612 into isolation chamber 630, as shown in FIG. 7B.

[0253] The initial volume of body fluid can be any suitable volume of body fluid, such as either the volume or amount described above. In some implementations, when the initial volume of body fluid is transferred into the isolation chamber 630, the flow of the body fluid can be stopped and / or the pressure difference can be substantially equalized, whereby the flow of the body fluid can be decelerated or stopped. In such implementations, the actuator 650 can then transition from its second state to its third state. In other implementations, the transition of the actuator 650 through the three states can be a substantially continuous transition. In such implementations, the initial volume of body fluid can be the volume of body fluid transferred into the isolation chamber 630 when the actuator 650 transitions from its first state to its second state, and continuing to transition the actuator 650 from its second state to its third state can act to stop the flow into the isolation chamber 630.

[0254] The actuator 650 can transition from its second state to its third state when an initial volume of body fluid is contained within the isolation portion 630. As shown in FIG. 7C, transitioning the actuator 650 to the third state can include transitioning and / or moving the second member 660 relative to the inlet 612 and the first member 651 of the actuator 650. Transitioning and / or moving the second member 660 causes the first end of the second member to transition and / or move from the second side of the inlet 612 to the first side of the inlet 612, thereby isolating and / or fluidly separating the isolation chamber 630 from the inlet 612. Further, the transition and / or movement of the second member 660 relative to the first member 651 can reduce the volume of the portion of the isolation chamber 630 disposed therebetween. In some implementations, the reduction in the volume of a portion of the isolation chamber 630 can result in an increase in pressure that acts to transition the valve 653 from a closed state to an open state, thereby enabling at least a portion of the initial volume of body fluid to be transferred into the sampling portion 635 of the isolation chamber 630, as shown in FIG. 7C.

[0255] As shown in FIG. 7D, the rapid test device 670 can be coupled to the housing 610 and / or placed in fluid communication with the sampling portion 635 of the isolation chamber 630 in some other way (e.g., via the coupling member 678). Thus, at least a portion of the body fluid can be transferred from the sampling portion 635 of the isolation chamber 630 into the rapid test device 670. In some implementations, the transfer of a portion of the initial volume can be automatic. In other implementations, the transfer of a portion of the initial volume can be responsive to one or more user inputs (e.g., via the actuator 650 and / or any other suitable actuation mechanism not shown in FIGS. 7A-7D). In some embodiments, by transferring a portion of the initial volume of the body fluid to the rapid test device 670, testing and / or analysis of a portion of the initial volume of the body fluid can be initiated as described in detail above with reference to the rapid test device 270. Although not shown in FIGS. 7A-7D, in some examples, the system 600, the transfer device 605, and / or the rapid test device 670 can be configured to provide a buffer (or any other suitable solution) that can be mixed with a portion of the initial volume of the body fluid. The rapid test device 670 can be configured to perform any suitable test and / or assay. For example, the rapid test device 670 can be an LFA configured to test for the presence of lactate, IL6, PCT, CD64, and / or NNM as described in detail above. Further, when the test or assay is complete, the rapid test device 670 can be configured to output test results that can be detected and / or determined by a human and / or one or more electronic devices as described in detail above with reference to the rapid test devices 170, 270, 370, and / or 470.

[0256] As described above, by transitioning the actuator 650 from its second state to its third state, a body fluid of an initial volume can be isolated, separated, and / or retained within the isolation chamber 630 and / or the rapid test device 670, and thus contaminants in the initial volume can also be isolated. Further, the arrangement of the rapid test device 670 can be such that the tests and / or assays performed by the rapid test device 670 are not affected by such contamination, which, as described in detail above, means that the accuracy of the test results output by the rapid test device 670 is not affected by such contamination.

[0257] As shown in FIGS. 7C and 7D, by transitioning the actuator 650 from its second state to its third state, fluid communication is established between the inlet 612 and the outlet 613 via the fluid flow path 615 disposed between the first end and the second end of the second member 660 of the actuator 650. More specifically, when the actuator 650 is in its third state, the first end of the second member 660 is disposed on the first side of the inlet 612, and the second end of the second member 660 is disposed on the second side of the outlet 613. In other words, both the inlet 612 and the outlet 613 are disposed between the first end and the second end of the second member 660. Thus, the fluid flow path 615 can establish fluid communication between the inlet 612 and the outlet 613 when the actuator 650 is in its third state.

[0258] In some implementations, the outlet 613 can be placed in fluid communication with a fluid collection device (not shown in FIGS. 7A-7D) before or after the actuator 650 is placed in its third state. As described in detail above, the fluid collection device can define a negative pressure and / or suction force that can act to draw body fluid into the fluid collection device and / or can be configured to generate such negative pressure and / or suction force. Thus, in response to the negative pressure and / or suction force, one or more subsequent volumes of body fluid can flow from the inlet 612, through the fluid flow path 615, through the outlet 613, and into the fluid collection device. Thus, as described above, by isolating an initial volume of body fluid within the isolation chamber 630 prior to collecting or obtaining one or more sample volumes of body fluid, the amount of contaminants in the one or more sample volumes is reduced and / or substantially eliminated. Thus, the system 600 can obtain an initial volume of body fluid for use in rapid tests that have a relatively low sensitivity to contamination and a subsequent volume of body fluid for use in tests that have a relatively high sensitivity to contamination, as described above with reference to systems 100, 200, 300, and / or 400.

[0259] FIGS. 8 and 9A-9D illustrate a fluid transfer and assay system 700 according to one embodiment. The fluid transfer and assay system 700 (also referred to herein as the "system") can include at least a fluid transfer device 705 and a rapid diagnostic test device 770. Some portions and / or aspects of the fluid transfer device 705 and / or the rapid diagnostic test device 770 can be the same as and / or substantially the same as the fluid transfer devices 105, 205, 305, 405, 505, and / or 605 and / or the rapid diagnostic test devices 170 (and / or LFA 170A, 170B), 270, 370, 470, 570, and / or 670 described in detail above, respectively. Thus, such some portions and / or aspects are not described in further detail herein.

[0260] The fluid transfer device 705 (also referred to herein as the "transfer device") can be of any suitable shape, size, and / or configuration. In some implementations, the transfer device 705 can be configured to draw body fluid (e.g., blood) from a patient into the transfer device 705 and / or through the transfer device 705. Additionally, the transfer device 705 can transfer at least a portion of the drawn body fluid to one or more other devices, such as the rapid diagnostic test device 770 and / or one or more fluid collection devices (not shown in FIGS. 8 and 9A - 9D), reservoirs, containers, vials, machines, tests, assays, etc.

[0261] The transfer device 705 includes at least a housing 710 and an actuator 750. The housing 710 of the device 705 can be of any suitable shape, size, and / or configuration. For example, in some embodiments, the housing 710 can be similar to and / or substantially the same as at least the housing 610 described above. Specifically, the housing 710 can have and / or form an inlet 712 and an outlet 713, and can define a fluid flow path 715 between the inlet 712 and the outlet 713. The inlet 712 can be any suitable inlet or port and can be configured to establish fluid communication between the housing 710 and a body fluid source (e.g., a patient). The outlet 713 can be any suitable outlet or port and can be configured to establish fluid communication between the housing 710 and a fluid collection device (not shown in FIGS. 8 - 9D), such as any of the fluid collection devices described in detail above. The fluid flow path 715, at least partially defined by the housing 710, extends between the inlet 712 and the outlet 713 and can selectively establish fluid communication between the inlet 712 and the outlet 713, as described in more detail herein.

[0262] As described above with reference to at least housing 610, housing 710 shown in FIGS. 8 - 9D includes, forms, and / or is connected to an isolation chamber 730 configured to be selectively placed in fluid communication with a fluid flow path and / or at least an inlet 712. Additionally, the housing defines an opening 721 and / or ports configured to receive a portion of rapid diagnostic test device 770, as described in more detail herein. Isolation chamber 730 can have any suitable shape, size, and / or configuration. For example, in some embodiments, isolation chamber 730 and / or at least a portion thereof can be substantially similar to isolation chambers 330, 430, and / or 630 described in detail above, at least in form and / or function. Accordingly, some portions and / or aspects of isolation chamber 730 are not described in further detail herein.

[0263] Actuator 750 of device 705 can be of any suitable shape, size, and / or configuration. In some embodiments, actuator 750 and / or aspects or portions thereof can be similar to and / or substantially the same as actuators 150, 250, 350, 450, and / or 650 described in detail above. In some embodiments, actuator 750 can be at least partially disposed within and / or partially formed by housing 710. As described above, actuator 750 can be configured to control, direct, and / or otherwise facilitate a selective flow of fluid through at least a portion of housing 710 and / or at least a portion of one or more fluid flow paths. Actuator 750 can be any suitable member or device configured to transition between any number of states (e.g., two, three, four, or more) in any suitable manner (e.g., user - actuated, auto - actuated, mechanically actuated, electronically actuated, chemically actuated, etc.).

[0264] More specifically, as shown in FIGS. 9A-9D, the actuator 750 includes a first member 751, a second member 760, and a third portion 765. The first member 751 of the actuator 750 can be of any suitable shape, size, and / or configuration. For example, the first member 751 can be similar to the first member 651 of the actuator 650 described in detail above, at least in form and / or function. The first member 751 includes at least one seal 752 disposed at a first end of the first member 751. The arrangement of the seals 752 can be such that the seals 752 engage and / or contact the inner surface of the housing 710 to form and / or define a substantial fluid seal between the seals 752 and the housing 710.

[0265] The first end of the first member 751 also includes a port 725 that is in fluid communication with the sampling channel 735. In some embodiments, for example, the port 725, as described above with reference to port 625, can be (i) configured to release and / or permit a flow of air or gas from the sampling channel 735 and (ii) a valve, coupler, and / or any suitable reconfigurable member or device configured to receive a portion of the rapid inspection device 770 and place the rapid inspection device 770 in fluid communication with the sampling channel 735. The sampling channel 735 is disposed within and / or defined by the first member 751. For example, in some embodiments, the first member 751 can have a hollow elongated portion that defines the sampling channel 735. Further, such a portion of the first member 751 can define and / or include openings, ports, valves, selectively permeable members, etc. configured to selectively place the sampling channel 735 in fluid communication with the isolation chamber 730. In some embodiments, the sampling channel 735 is included in and / or defined by the first member 751 of the actuator 750, but the sampling channel 735 can be similar to the sampling portion 635 of the isolation chamber 630 described above with reference to FIGS. 7A-7D, at least in form and / or function.

[0266] As shown in FIGS. 9A-9D, the first member 751 also includes an engagement member 755 disposed at or on a second end of the first member 751 opposite the first end. The engagement member 755 can be of any suitable shape, size, and / or configuration. For example, in some embodiments, the engagement member 755 can be a protrusion, tab, button, knob, and / or any other suitable engagement member. The engagement member 755 is configured to selectively engage with a portion of a third member 765 of the actuator 750 to induce and / or at least partially control relative movement between the first member 751, the second member 760, and / or the third member 765, as will be described in more detail herein.

[0267] The second member 760 of the actuator 750 can be of any suitable shape, size, and / or configuration. As shown in FIGS. 9A-9D, the second member 760 can be disposed around and / or on at least a portion of the first member 751. The second member 760 includes a set of seals 761. As shown, the second member 760 can include a first end having an inner seal 761 and an outer seal 761, and a second end opposite the first end having an outer seal 761. In this way, the second member 760 can be similar to and / or substantially the same as the second member 660 of the actuator 650. Accordingly, the second member 760 and / or aspects or portions thereof will not be described in further detail herein.

[0268] The third member 765 can be of any suitable shape, size, and / or configuration. In some embodiments, the third member 765 can be included in and / or can form a part of the housing 710 and / or an outer portion of the transfer device 705. For example, as shown in FIGS. 9A-9D, at least a portion of the housing 710, the first member 751, and the second member 760 can be disposed within a part of the third member 765. More particularly, the third member 765 can be, for example, a substantially hollow cylinder having an open end and a substantially closed end. The substantially closed end includes and / or defines a detent, a recess, an opening, and / or an engagement structure (referred to herein as "engagement structure 766"). The engagement structure 766 can contact and / or otherwise selectively engage an engagement member 755 of the first member 751. For example, as described in more detail herein, the engagement member 755 can be configured to engage and / or contact the engagement structure 766, whereby the first member 751 and the third member 765 can be moved collectively and / or simultaneously when the actuator 750 is transitioned between two or more states or configurations. Further, a portion of the transition of the actuator 750 can disengage and / or move the engagement member 755 relative to the engagement structure 766, whereby the first member 751 can be moved relative to the third member 765 (or the third member 765 can be moved relative to the first member 751) as described in more detail herein.

[0269] As shown in FIGS. 9A-9D, the arrangement of the first member 751 and the second member 760 of the actuator is such that an isolation chamber 730 is arranged and / or defined, for example, between a first end of the first member 751 and a first end of the second member 760. Additionally, the second member 760 is configured to at least partially define a fluid flow path 715 between a first end and a second end of the second member 760. Accordingly, the first end of the second member 760 and a seal 761 included in the first end isolate and / or fluidly isolate the isolation chamber 730 from the fluid flow path 715.

[0270] The actuator 750 is configured to transition between at least a first state, a second state, a third state, and a fourth state. As shown in FIGS. 9A-9D, the first end of the first member 751 and the seal 752 included therein are disposed and maintained on the first side of the inlet 712 and the first side of the outlet 713 regardless of the state of the actuator 750. Similarly, the second end of the second member 760 and the seal member 761 included therein are disposed and maintained on the second side of the inlet 712 (opposite the first side) and the second side of the outlet 713 (opposite the first side) regardless of the state of the actuator 750. However, the first end of the second member 760 and the seal member 761 disposed thereon are (i) disposed on the second side of the inlet 712 and the first side of the outlet 713 when the actuator 750 is in the first state (FIG. 9A), the second state (FIG. 9B), and the third state (FIG. 9C), and (ii) configured to be disposed on the first side of the inlet 712 and the first side of the outlet 713 when the actuator 750 is in the fourth state (FIG. 9D). Thus, the arrangement of the actuator 750 is such that, as described in more detail herein, the transition of the actuator 750 can selectively direct and / or divert the flow of fluid between (i) the inlet 712 and the isolation chamber 730 and (ii) the inlet 712 and the outlet 713 via the fluid flow path 715.

[0271] The rapid diagnostic test device 770 (also referred to herein as the "rapid test device" or simply the "test device") can be any suitable test device. For example, the rapid test device 770 can be an LFA as described in detail above with reference to the LFA170A shown in FIG. 2A or the LFA170B shown in FIG. 2B. In some implementations, the test device 770 and / or aspects or portions thereof can be substantially similar to the rapid test devices 170, 270, 370, 470, 570, and / or 670 described in detail above. Thus, the transfer device 770 and / or aspects or portions thereof are not described in further detail herein.

[0272] As shown in FIG. 9D, the rapid inspection device 770 includes a connecting member 778 that is connected to and / or at least in fluid communication with the substrate 771 of the inspection device 770 (e.g., directly connected to the substrate 771 and / or connected via a connecting mechanism such as the mounting mechanism 579). The connecting member 778 is at least partially inserted through the opening 721 of the housing 710 to establish fluid communication with the sampling channel 735 when the rapid inspection device 770 is connected to the transfer device 705. For example, the connecting member 778 can be a piercing member, a needle, a tube, a capillary, etc. that can pierce the port 725 and / or advance through the port 725 in other ways. In some embodiments, the substrate 771 and the connecting member 778 can be substantially similar to the substrates 571 and / or 671, and the connecting members 578 and / or 678 described in detail above. Therefore, the substrate 771 and the connecting member 778 (and / or their aspects or parts) will not be described in further detail herein.

[0273] The system 700 can be used to obtain one or more volumes of body fluid from a patient for use in one or more tests, assays, and / or diagnostic methods. As described above, for example, the inlet 712 can be placed in fluid communication with a body fluid source. The actuator 750 can be in a first state when the inlet 712 is placed in fluid communication with a body fluid source (e.g., a part of a patient), thereby establishing fluid communication between the inlet 712 and the isolation chamber 730 and isolating the outlet 713 from the inlet 712, as shown in FIG. 9A. Further, when the actuator 750 is in the first state, the first end of the first member 751 can be near or adjacent to the first side of the inlet 712, and the first end of the second member 760 can be near or adjacent to the second side of the inlet 712. In this way, the isolation chamber 730 defined between the first member 751 and the second member 760 can have a first volume.

[0274] In some examples, when the inlet 712 is placed in fluid communication with the body fluid source, the actuator 750 can transition from its first state to its second state. For example, as shown in FIG. 9B, the user can apply a force to the third member 765 to act to move the third member 765 relative to the housing 710. As described above, the arrangement of the engaging member 755 of the first member 751 and the engaging structure 766 of the third member 765 is such that when the third member 765 moves relative to the housing 710, the first member 751 moves similarly. The movement of the first member 751 is also performed relative to the second member 760 (for example, the second member 760 has not yet moved), thereby increasing the volume of the isolation chamber 730 disposed between the first member 751 and the second member 760. In addition, the transition and / or movement of the first member 751 can reduce the volume within the housing 710 on the side of the first member 751 opposite the isolation chamber 730, and the opening 721 can be an opening that allows air or gas contained therein to leak and / or flow out from the sampling channel 735. Thus, when the actuator 750 transitions from its first state (FIG. 9A) to its second state (FIG. 9B), a negative pressure difference can be generated within the isolation chamber that can act to draw an initial volume of body fluid from the body fluid source through the inlet 712 into the isolation chamber 730, as described in detail above with reference to the isolation chamber 630. Further, the initial volume of body fluid can be any suitable volume of body fluid, such as any of the volumes or amounts described above.

[0275] Actuator 750 can transition from its second state (FIG. 9B) to its third state (FIG. 9C) when an initial volume of body fluid is contained within isolation 730. As described above with reference to transfer device 605, the transition of actuator 750 from its second state to its third state can occur in response to an initial volume of body fluid being placed within isolation chamber 730, one or more pressure differences being equalized, and at a given point in the continuous process, actuator 750 transitioning from its first state to its fourth state, among other things. In some examples, the transition can occur automatically or in response to an applied force.

[0276] As shown in FIG. 9C, transitioning actuator 750 to its third state can include transitioning and / or moving first member 751 and third member 765 an additional amount relative to housing 710 and second member 760. More specifically, first member 751, when in its third state, can be positioned relative to second member 760 such that an opening, port, valve, etc. (referred to herein as "opening 754") is in fluid communication with isolation chamber 730 and / or inlet 712, as shown in FIG. 9C. In this way, a volume of body fluid within sampling channel 735 defined by first member 751 can be transferred. As described above, in some embodiments, port 725 can be configured to vent sampling channel 735 to facilitate the flow of body fluid into sampling channel 735.

[0277] With a volume of body fluid contained within sampling channel 735, actuator 750 can transition from its third state (FIG. 9C) to its fourth state (FIG. 9D). More specifically, in some implementations, third member 765 and second member 760 can be moved relative to housing 710, while first member 751 is maintained in a position substantially fixed relative to housing 710. Stated another way, third member 765 and second member 760 are moved together and relative to first member 751.

[0278] As shown in FIG. 9D, when the actuator 750 transitions to the fourth state, the engagement member 755 is disengaged from the engagement surface 766 and / or is moved relative to the engagement surface 766. In some embodiments, the engagement member 755 and / or the engagement surface 766 may be sized and / or configured to maintain contact and / or engagement until a desired and / or predetermined force sufficient to overcome the force maintaining the engagement (e.g., frictional force, a force sufficient to elastically and / or plastically deform the engagement member 755 and / or the engagement surface 766, and / or any other suitable force) is applied. In other words, the third member 765 may be moved relative to the first member 751 when the force meets a criterion and / or is greater than a threshold amount of force.

[0279] The second member 760 of the actuator 750 is moved in the same direction as the third member 765 when the actuator 750 transitions to the fourth state. As shown in FIG. 9D, when the second member 760 transitions and / or moves, the first end of the second member 760 transitions and / or moves from the second side of the inlet 712 to the first side of the inlet 712, thereby isolating and / or fluidly separating the isolation chamber 730 from the inlet 712. Further, when the second member 760 transitions and / or moves relative to the first member 751, the opening 754 of the first member 751 can be placed opposite the inner seal 561 contained in or on the first end of the second member 760, thereby, in some examples, enabling the sampling channel 735 to be vented, as described in more detail herein.

[0280] As shown in FIG. 9D, the rapid test device 770 is coupled to the housing 710 and / or, in other cases, at least partially inserted into and / or through the opening 721 of the housing 710 such that the connecting member 778 can establish fluid communication with the sampling channel 735 (e.g., via port 725). Thus, at least a portion of the body fluid can be transferred from the sampling channel 735 into the rapid test device 770 as described in detail above with reference to the rapid test devices 470, 570, and / or 670. In some embodiments, by transferring a volume of body fluid from the sampling channel 735 into the rapid test device 770 as described in detail above with reference to the rapid test device 270, testing and / or assay of a portion of the initial volume of body fluid and / or testing and / or assay regarding a portion of the initial volume of body fluid can be initiated. Further, in some examples, by venting the sampling channel 735 through the opening 754, a desired pressure differential within the sampling channel 735 can be enabled that facilitates the transfer of body fluid from the sampling channel 735 into the rapid test device 770. The rapid test device 770 can be configured to perform any suitable test and / or assay, such as any of the tests and / or assays described in detail above (e.g., tests regarding the presence of lactate, IL6, PCT, CD64, and / or NNM). Further, upon completion of the test or assay, the rapid test device 770 can be configured to output test results that can be detected and / or determined by a human and / or one or more electronic devices as described in detail above with reference to the rapid test devices 170, 270, 370, 470, 570, and / or 670.

[0281] As described above, by transitioning the actuator 750 from its third state to its fourth state, the initial volume of body fluid within the isolation chamber 730 and / or the rapid test device 770 can be isolated, separated, and / or retained, and thus contaminants within the initial volume can also be isolated. Further, the array configuration of the rapid test device 770 can be such that the tests and / or assays performed by the rapid test device 770 are not affected by such contamination, which, as described in detail above, means that the accuracy of the test results output by the rapid test device 770 is not affected by such contamination.

[0282] As shown in FIG. 9D, by transitioning the actuator 750 from its third state to its fourth state, fluid communication is established between the inlet 712 and the outlet 713 via the fluid flow path 715 disposed between the first and second ends of the second member 760 of the actuator 750. As described in detail above with reference to the actuator 650, when the actuator 750 is in its fourth state, the first end of the second member 760 is disposed on the first side of the inlet 712, and the second end of the second member 760 is disposed on the second side of the outlet 713.

[0283] In some implementations, the outlet 713 can be placed in fluid communication with a fluid collection device (not shown in FIGS. 8-9D) before or after the actuator 750 is placed in its fourth state. As described in detail above, the fluid collection device can define a negative pressure and / or suction force that can act to draw body fluid into the fluid collection device, and / or can be configured to generate such negative pressure and / or suction force. Thus, in response to the negative pressure and / or suction force, one or more subsequent volumes of body fluid can flow from the inlet 712, through the fluid flow path 715, through the outlet 713, and into the fluid collection device. Thus, as described above, by isolating an initial volume of body fluid within the isolation chamber 730 before collecting or obtaining one or more subsequent volumes of body fluid, the amount of contaminants in the one or more subsequent volumes is reduced and / or substantially eliminated. Thus, the system 700 can obtain an initial volume of body fluid for use in rapid tests that are relatively insensitive to contamination and a subsequent volume of body fluid for use in tests that are relatively sensitive to contamination, as described above with reference to systems 100, 200, 300, 400, and / or 600.

[0284] FIGS. 10, 11, and 12A-12D illustrate a fluid transfer and assay system 800 according to one embodiment. The fluid transfer and assay system 800 (also referred to herein as the "system") can include at least a fluid transfer device 805 and a rapid diagnostic test device 870. Some parts and / or aspects of the fluid transfer device 805 and / or the rapid diagnostic test device 870 can be the same as and / or substantially the same as the fluid transfer devices 105, 205, 305, 405, 505, 605, and / or 705, and / or the rapid diagnostic test devices 170 (and / or LFA 170A, 170B), 270, 370, 470, 570, 670, and / or 770 described in detail above. Thus, some such parts and / or aspects are not described in further detail herein.

[0285] The fluid transfer device 805 (also referred to herein as the "transfer device") can be of any suitable shape, size, and / or configuration. In some implementations, the transfer device 805 can be configured to draw body fluid (e.g., blood) from a patient into and / or through the transf...

Claims

**Claim 1** A system for the early detection and treatment of sepsis, comprising a fluid transfer device having an inlet and an outlet, the inlet being configured to receive a flow of body fluid from a body fluid source; and at least one flow-based assay device configured to be coupled to the fluid transfer device, wherein a portion of the at least one flow-based assay device engages the outlet when coupled to the fluid transfer device to enable transfer of a first volume of a portion of the body fluid from the fluid transfer device to the at least one flow-based assay device, and the at least one flow-based assay device is configured to detect at least one sepsis-related biomarker. **Claim 2** The system of claim 1, wherein the at least one flow-based assay device is one of a sandwich lateral flow assay device or a competitive lateral flow assay device. **Claim 3** The at least one flow-based assay device comprises a conjugate element comprising a labeled bioactive agent configured to bind to the at least one sepsis-related biomarker; and one or more capture elements configured to immobilize each of the at least one sepsis-related biomarker and the labeled bioactive agent, wherein accumulation of the labeled bioactive agent immobilized along the one or more capture elements provides a visual indicator associated with the presence of each of the at least one sepsis-related biomarker in the portion of the first volume of body fluid. **Claim 4** The system of claim 3, wherein the labeled bioactive agent comprises at least one of an antibody, an aptamer, and a protein binder. **Claim 5** The system of claim 1, wherein the fluid is blood. **Claim 6** The system of claim 1, wherein the at least one sepsis-related biomarker is at least a portion of one of procalcitonin, lactate, surface antigen classification 64, neutrophil count marker, and interleukin 6. **Claim 7** The system of claim 6, wherein the neutrophil count marker is one of neutrophil elastase, lactoferrin, myeloperoxidase, and human neutrophil lipocalin.

8. The system according to claim 1, wherein the at least one sepsis-related biomarker is at least a part of surface antigen classification 64 and the neutrophil count biomarker.

9. A system for early detection and treatment of sepsis, a fluid transfer device having an inlet and an outlet, the inlet being configured to receive a flow of body fluid from a body fluid source, at least one flow-based assay device configured to be coupled to the fluid transfer device, a processing circuit, and a part of the at least one flow-based assay device engages with the outlet when coupled to the fluid transfer device, enabling a first volume of a part of the body fluid to be transferred from the fluid transfer device to the at least one flow-based assay device, the at least one flow-based assay device being configured to detect at least one sepsis-related biomarker, the processing circuit receives data corresponding to the at least one sepsis-related biomarker from the at least one flow-based assay device, applies a computational model to the received data, generates a sepsis probability score based on an output of the applied computational model, and is configured to alert a healthcare provider to initiate a corresponding treatment when the sepsis probability score exceeds a sepsis-related threshold.

10. The system according to claim 9, wherein the at least one flow-based assay device is one of a sandwich lateral flow assay device or a competitive lateral flow assay device.

11. The at least one flow-based assay device includes a conjugate element including a labeled bioactive agent configured to bind to the at least one sepsis-related biomarker, and one or more capture elements configured to immobilize each of the at least one biomarker related to sepsis and the labeled bioactive agent, wherein an accumulation of the labeled bioactive agent immobilized along the one or more capture elements provides a visual indicator associated with the presence of each of the at least one sepsis-related biomarker in the first volume of the body fluid.

12. The system of claim 11, wherein the labeled bioactive agent comprises at least one of an antibody, an aptamer, and a protein binder. **Claim 13** The system of claim 9, wherein the fluid is blood. **Claim 14** The system of claim 9, wherein the at least one sepsis-related biomarker is at least a part of one of procalcitonin, lactate, surface antigen classification 64, neutrophil count marker, and interleukin 6. **Claim 15** The system of claim 14, wherein the neutrophil count marker is one of neutrophil elastase, lactoferrin, myeloperoxidase, and human neutrophil lipocalin. **Claim 16** The system of claim 9, wherein the at least one sepsis-related biomarker is at least a part of surface antigen classification 64 and the neutrophil count marker. **Claim 17** The system of claim 9, wherein the computational model is based on a random forest classifier. **Claim 18** The system of claim 9, wherein the computational model is trained on a reference dataset comprising at least one of point-of-care metrics and historical metrics. **Claim 19** The system of claim 18, wherein the point-of-care metrics comprise one or more of lactate, interleukin 6, surface antigen classification 64, procalcitonin, neutrophil count marker, heart rate, blood pressure, white blood cell count, respiratory rate, and body temperature. **Claim 20** The system of claim 19, wherein the neutrophil count marker is one of neutrophil elastase, lactoferrin, myeloperoxidase, and human neutrophil lipocalin. **Claim 21** The system of claim 18, wherein the historical metrics can be derived from one or more of medical history data, previous diagnoses, treatment plans and medications, laboratory and test results, immunization details and dates, and medical images. **Claim 22** A method for early detection and treatment of sepsis, comprising: receiving data associated with at least one flow-based assay device corresponding to at least one sepsis-related biomarker; applying a computational model to the received data; generating a sepsis probability score based on the output of the applied computational model; A method comprising warning a healthcare provider to initiate a corresponding treatment when the sepsis probability score exceeds a sepsis-related threshold.

23. The method according to claim 22, wherein the at least one flow-based assay device is one of a sandwich lateral flow assay device or a competitive lateral flow assay device.

24. The method according to claim 22, wherein the at least one sepsis-related biomarker is at least a part of one of procalcitonin, lactate, surface antigen classification 64, neutrophil count marker, and interleukin 6.

25. The method according to claim 24, wherein the neutrophil count marker is one of neutrophil elastase, lactoferrin, myeloperoxidase, and human neutrophil lipocalin.

26. The method according to claim 22, wherein the at least one sepsis-related biomarker is at least a part of surface antigen classification 64 and the neutrophil count marker.

27. The method according to claim 22, wherein the computational model is based on a random forest classifier.

28. The method according to claim 22, wherein the computational model is trained on a reference dataset including at least one of point-of-care metrics and historical metrics.

29. The method according to claim 28, wherein the point-of-care metrics include one or more of lactate, interleukin 6, surface antigen classification 64, procalcitonin, neutrophil count marker, heart rate, blood pressure, white blood cell count, respiratory rate, and body temperature.

30. The method according to claim 29, wherein the neutrophil count marker is one of neutrophil elastase, lactoferrin, myeloperoxidase, and human neutrophil lipocalin.

31. The method according to claim 28, wherein the historical metrics can be derived from one or more of medical history data, previous diagnoses, treatment plans and medications, laboratory and test results, immunization details and dates, and medical images.

32. A method for the early detection and treatment of sepsis, comprising placing the inlet of a fluid transfer device in fluid communication with a body fluid source, and receiving body fluid into the fluid transfer device from the inlet. Establishing fluid communication between the inlet and outlet of the fluid transfer device to enable a volume of body fluid to flow into a sample reservoir in fluid communication with the outlet; Conveying a portion of the volume of body fluid to a sample element of at least one flow-based assay device that is at least temporarily fluidly connected to the fluid transfer device; Conveying a buffer solution to the sample element of the at least one flow-based assay device, the method comprising.

33. The at least one flow-based assay device is a lateral flow assay device, The method is, Performing a lateral flow assay on the portion of the volume of body fluid; Further comprising providing an output associated with the result of the lateral flow assay, the method according to claim 32.

34. The lateral flow assay device is one of a sandwich lateral flow assay device or a competitive lateral flow assay device, the method according to claim 33.

35. The lateral flow assay device is, A conjugate element comprising a labeled bioactive agent configured to bind to each target analyte; A capture element configured to immobilize each of the target analytes and the labeled bioactive agent bound thereto, Accumulation of the labeled bioactive agent immobilized along the capture element is configured to provide an indicator associated with the presence of each target analyte in the first volume of the portion of the body fluid, the method according to claim 33.

36. The bioactive agent comprises at least one of an antibody, an aptamer, and a protein binder, the method according to claim 35.

37. The lateral flow assay device is, A conjugate element comprising a labeled antibody configured to bind to at least a portion of one of procalcitonin, surface antigen classification 64, neutrophil count marker, interleukin 6, or lactate, respectively; At least one capture element configured to immobilize at least a portion of one or more of procalcitonin, surface antigen classification 64, the neutrophil count marker, interleukin 6, or lactate, respectively, and the labeled antibody bound thereto, The accumulation of the labeled antibodies immobilized along the at least one capture element is configured to provide an indicator associated with the presence of one or more of procalcitonin, surface antigen classification 64, the neutrophil count marker, interleukin 6, or lactate in the portion of the first volume of body fluid, respectively. The method according to claim 33.

38. The method according to claim 37, wherein the neutrophil count marker is one of neutrophil elastase, lactoferrin, myeloperoxidase, and human neutrophil lipocalin.

39. The method according to claim 33, further comprising outputting the results of the lateral flow assay device to an electronic device configured to predict the likelihood that each patient has sepsis.

40. The method according to claim 32, further comprising lysing monocytes within the portion of the volume of body fluid transported to the sample element.

41. The method according to claim 32, further comprising lysing neutrophils within the portion of the volume of body fluid transported to the sample element.

42. The method according to claim 37, further comprising cleaving the extracellular domain of surface antigen classification 64 from neutrophils within the portion of the volume of body fluid transported to the sample element.

43. The method according to claim 42, wherein the extracellular domain of surface antigen classification 64 is cleaved by a proteolytic peptidase.

44. The method according to claim 43, wherein the proteolytic peptidase is an endopeptidase.

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