Method and device for processing at least one body fluid sample for pathogenic microorganism analysis

Abstract

The invention relates to a method and device for processing at least one body fluid sample for pathogenic microorganism analysis. The method includes the steps of providing at least one body fluid sample, filtering the at least one body fluid sample through at least one catch filter which is configured for capturing at least a fraction of pathogenic microorganisms from the at least one body fluid sample if present in the body fluid sample, and applying at least one stress stimulus to at least part of the at least one catch filter, in particular to the fraction of pathogenic microorganisms, such that at least a fraction of the captured pathogenic microorganisms extracellularly secrete stress responsive biomolecules.

Description

[0001] Method and device for processing at least one body fluid sample for pathogenic microorganism analysis

[0002] The invention relates to a method for processing at least one body fluid sample for pathogenic microorganism analysis. The invention also relates to a device for processing at least one body fluid sample for pathogenic microorganism analysis.

[0003] Current diagnostics processes for pathogenic microorganism detection and identification thereof are based mainly on phenotypic methods or molecular diagnostics methods. Examples of phenotypic methods for, for example, bacterial identification can be based on protein expression, or ‘fingerprints’ whereas molecular diagnostics methods are typically based on for example genomic markers and / or nucleic acid sequences. However, the instrumentation and equipment and consumables of these diagnostic tools are rather expensive and the involved diagnostic methods are time consuming. Additionally, professionally trained people are required to execute these tests and most of these tools are not used in the point-of-care setting. Further, not all clinical sample types can be processed with every diagnostic tool.

[0004] Hence, a goal of the invention is to provide a more simplified solution to perform fluid analysis, in particular body fluid analysis, in an more time efficient manner.

[0005] The invention provides thereto a method for processing at least one fluid sample for analyte analysis, comprising the steps of:

[0006] A) providing at least one fluid sample,

[0007] B) isolating at least a fraction of at least one analyte from the at least one fluid sample if present said fluid sample, and

[0008] C) subjecting at least part of the fraction of at least one analyte, to at least one environmental change such that at least a fraction of the captured analyte generates an environmental response.

[0009] Preferably, the fluid sample is at least one body fluid sample, such as blood, urine, sweat, and the like. The analyte to be analysed may be viable cell(s). Viable cell(s) may be any type of cell, such as prokaryotic cell(s) and / or eukaryotic cell(s). The step of isolating at least a fraction of at least one analyte may be performed by filtering, centrifugation, flow cytometry, magnetic bead extraction, and / or combinations thereof. The environment of the analyte may be changed by applying a stimulus to at least a part of the fraction of at least one analyte and / or to the fluid sample.

[0010] A preferred application of the method and device according to the present invention is related to processing at least one body fluid sample for pathogenic microorganism or analysis.

[0011] The invention provides thereto in a preferred embodiment a method for processing at least one body fluid sample for pathogenic microorganism analysis, comprising the steps of:

[0012] A) providing at least one body fluid sample,

[0013] B) isolating at least a fraction of pathogenic microorganisms from the at least one body fluid sample, if present in the body fluid sample, in particular by filtering the at least one body fluid sample through at least one catch filter which is configured for capturing at least a fraction of pathogenic microorganisms from the at least one body fluid sample if present in the body fluid sample, and

[0014] C) applying at least one stimulus, in particular at least one stress stimulus to at least part of the at least one catch filter and / or to the fraction of pathogenic microorganisms, if captured, in particular such that at least a fraction of the captured pathogenic microorganisms extracellularly secrete and / or excrete stress responsive biomolecules.

[0015] The method according to the present invention enables that a body fluid sample can be prepared for pathogenic microorganism analysis in an effective and efficient manner. The method thereby enables that a body fluid sample can be directly prepared for the analysis without requiring a cultivation or growth step. This results in that the processing time between the provision of the body fluid sample and the actual analysis thereof can be significantly reduced. Filtering of the at least one body fluid sample through at least one catch filter results in that at least part of any pathogenic microorganisms, if present in the body fluid, will be retained within the catch filter. The at least one catch filter is in particular configured for capturing at least a fraction of at least one pathogenic microorganism, and in particular for capturing multiple pathogenic microorganisms. The mesh size of at least one catch filter could be specifically chosen for a target pathogenic microorganism. Once at least part of at least one pathogenic microorganism is captured within the at least one catch filter, it was experimentally found that subjecting at least part of the captured pathogenic microorganism, directly or indirectly, to at least one (stress) stimulus causes the pathogenic microorganism to secrete and / or excrete stress responsive biomolecules. Therefore, the method according to the present invention includes the step of applying at least one stress stimulus to at least part of the at least one catch filter and / or to at least part of the fraction of captured pathogenic microorganisms, in particular such that at least a fraction of the captured pathogenic microorganisms extracellularly secretes and / or excretes stress responsive biomolecules. Hence, within the context of the present invention step C) could also be the step of applying (directly and / or indirectly) at least one stress stimulus to at least part of the fraction of pathogenic microorganisms. A benefit of this step is that the secreted and / or excreted stress responsive biomolecules can be more easily detected than the pathogenic microorganisms itself, in particular if no further preparation and / or cultivation steps have been applied. The presence of stress responsive biomolecules provides an indirect measure for the presence of pathogenic microorganisms in the body fluid. Subsequently, further treatment of the patient can be based upon the analysis results, and in particular upon the outcome whether or not pathogenic microorganisms are present. The method enables that diagnostics of body fluid sample can be done more time-efficiently which will have a positive effect on any further medical treatment, if needed, and the effectiveness thereof as treatment could be started in an earlier stage.

[0016] The method in particular focusses on direct processing of body fluid samples to test if pathogenic microorganisms are present. The method can be configured for use in combination with a device according to the present invention. Whilst the method could be applied for such, the method is not specifically configured to detection of pathogenic microorganisms which are spiked into a fluid sample.

[0017] The method benefits of being relatively simple, resulting in that the method does not require trained of qualified professionals to perform the processing of the body fluid sample according to the invention. The method could be performed in a manual way. However, it is not excluded that at least part of the method could be performed in an automated manner. Yet another benefit of the method according to the present invention is that the method only produces a relatively small amount of waste. The waste, typically the liquid waste, produced after the method is suitable for conventional medical disposal, in particular conventional biological disposal.

[0018] The method according to the invention is in particular suitable for direct testing of fluid samples, in particular body fluid samples. However, it is conceivable that preparation steps, such as prefiltration, purification and / or dilution of the (body fluid) sample, are done prior to step B) and / or step C). It is also not excluded to apply the method according to the present invention in combination with a cultured sample.

[0019] Within the context of the present invention, when it is referred to pathogenic microorganisms any type of pathogenic microorganisms could be meant. Nonlimiting example of pathogenic microorganisms are fungi and / or bacteria. When it is referred to a sample or a fluid sample, this could for example be a body fluid sample, more in particular a human and / or animal body fluid sample or a biological fluid, such as but not limited to saliva, blood, blood plasma, blood culture, urine, sweat, tears, cerebrospinal fluid, lymph fluid, synovial fluid, milk, amniotic fluid and / or derivatives thereof. Alternatively, the sample could be any further fluid that could contain at least one analyte, such as viable pathogens. Further non-limiting examples of fluid samples are water and / or solutions in the lab. The sample can be a pure sample. However, it is also possible that the sample is diluted and / or dissolved in a solvent. The viable pathogen could for example be bacteria, fungi, and / or parasites.

[0020] It is imaginable that the type of secreted and / or excreted stress responsive biomolecules depends on the type of stress applied to the fraction of pathogenic microorganisms. In particular, secretion and / or excretion of stress responsive biomolecules as response on the stress stimulus is substantially without cell disruption of the pathogenic microorganism(s). The pathogenic microorganisms may use different biological pathways for secretion and / or excretion of stress responsive biomolecules. The biological pathways may depend on the type of stress applied to the fraction of pathogenic microorganisms. Further analysis steps could be determined based on the applied embodiment. Filtering of the at least one body fluid sample through at least one catch filter such that at least part of any pathogenic microorganisms, if present in the body fluid sample, is retained within and / or on the catch filter is an example of isolating at least a fraction of pathogenic microorganisms from the at least one body fluid sample, if present in the body fluid sample. Alternatively step B) of isolating at least a fraction of pathogenic microorganisms from the at least one body fluid sample could be provided via centrifugation, flow cytometry, magnetic bead extraction and / or combinations thereof.

[0021] The method steps as applied in the method according to the present invention are in particular subsequent steps. Hence, in an embodiment, steps A) to C) are subsequent steps. The method enables that no growth or cultivation step of the pathogenic microorganisms, if captured, is needed but that instead at least one stress stimulus is applied which also enables accurate and effective analysis of any pathogenic microorganisms present in the body fluid sample. Hence, it is imaginable that directly after obtaining the body fluid sample an isolation step is performed. It is imaginable that steps A) to C) are performed in a time frame of between 30 seconds to 60 minutes, preferably within 1 minute to 30 minutes, more preferably within 5 minutes to 15 minutes, such as 10 minutes. This is possible, because no growth or cultivation step of the pathogenic microorganisms is needed. Therewith, the present invention provides a time efficient body fluid analysis.

[0022] Applying at least one stimulus, in particular at least one stress stimulus, to at least part of the at least one catch filter and / or to the fraction of pathogenic microorganisms, if captured, is an example of subjecting the at least part of the fraction of pathogenic microorganisms to at least one environmental change. Alternatively, an environmental change may be provided by changing the environment of the pathogenic microorganisms, for example by changing the temperature and / or the pH and / or the osmolality, and the like.

[0023] Pathogenic microorganisms typically extracellularly secrete and / or excrete stress responsive biomolecules as a response to the stress stimulus. Within the context of the present invention, stress is typically an environmental change which affects the captured pathogenic microorganisms. At least one stress stimulus could for example be a stress factor and / or a stress inducer. At least one stress stimulus may be an external stress stimulus or an internal stress stimulus. Hence, the purpose of applying at least one stress stimulus is triggering (extracellularly) secretion and / or excretion of stress responsive biomolecules. At least one stress stimulus could be applied directly and / or indirectly to at least part of the catch filter and / or to at least part of the captured pathogenic microorganisms. This could be done via several different approaches. At least one stress stimulus could for example be a chemical stress stimulus, a mechanical stress stimulus or an electrical stress stimulus. Also a combination of a chemical stress stimulus, a mechanical stress stimulus and an electrical stress stimulus could be applied. Hence, in a possible embodiment, during step C) at least one chemical stress stimulus is applied to at least part of the at least one catch filter. It is for example imaginable that at least one chemical stress stimulus is configured to the change the chemical environment of the captured pathogenic microorganisms. At least one chemical stress stimulus may for example create a change in pH, a change in the osmolarity, decrease the concentration of nutrients and / or cause starvation. It is imaginable that at least one stress stimulus is a fluid stress stimulus. In particular at least one chemical stress stimulus may be a fluid stress stimulus. At least one chemical stress stimulus may also comprise at least one fluid and / or at least one chemical substance, preferably chosen from the group consisting of: water, demiwater, an acid, a base, an antimicrobial substance and / or a combination thereof. Hence, in a possible embodiment, at least part of the at least one catch filter and / or at least part of the pathogenic microorganisms, if captured, is exposed to and / or brought into contact with at least one fluid and / or at least one chemical substance. It is also possible that step C) includes that at least part of the at least one catch filter is wetted. Step C) may include that that the pathogenic microorganisms are at least partially provided and / or dissolved in at least one fluid. In a beneficial embodiment, step C) includes that at least part of the at least one catch filter is brought into fluidic environment. Within the context of the present invention, the at least one fluid may also comprise at least one chemical substance and / or at least one chemical compound. The pathogenic microorganisms, if present, may be triggered to extracellular secrete and / or excrete stress responsive biomolecules.

[0024] In a possible embodiment, during step C) at least one mechanical stress stimulus is applied to at least part of the at least one catch filter. At least one mechanical stress stimulus can be configured to trigger extracellular activity of the captured pathogenic microorganisms. It is for example possible that at least one mechanical stress stimulus is configured to provide a mechanical modification to the pathogenic microorganisms and / or the environment of the pathogenic microorganisms. At least one mechanical stress stimulus could for example be configured to provide a mechanical modification of at least part of the at least one catch filter. This would result in a change of the environment of the captured pathogenic microorganisms, which could subsequently trigger the extracellular activity thereof. In a possible embodiment, at least one mechanical stress stimulus is a mechanical force. At least one mechanical stress stimulus could also involve or include at least one mechanical force. Non-limiting examples of mechanical force which could be applied are a compressive force, a tensile force, a shear force, or combinations thereof. It is imaginable that at least one mechanical force is applied to at least part of at least one catch filter. It is for example possible that at least part of at least one catch filter is stretched in at least one direction. This may cause a mechanical stress stimulus to at least part of the captured pathogenic microorganisms which are present upon said catch filter.

[0025] It is also possible that during step C) at least one electrical stress stimulus is applied to at least part of the at least one catch filter. In a possible embodiment, at least one electrical stress stimulus includes the provision of at least one electrical current. Applying an electrical current to at least part the catch filter could trigger extracellular activity of the pathogenic microorganisms captured by said catch filter.

[0026] In yet another possible embodiment, during step C) at least one stress stimulus is applied by providing a change in temperature. It is for example imaginable that a change in temperature of at least part of the catch filter is provided and / or a change in temperature of at least part of the captured pathogenic microorganisms.

[0027] Adjusting the temperature of at least part of the catch filter could result in a change of the temperature of the captured pathogenic microorganisms. It is for example possible that the step includes lowering or raising the temperature with at least 5 degrees Celsius, in particular at least 10 degrees Celsius, more in particular at least 20 degrees Celsius. It is also conceivable that during step C) at least one stress stimulus is applied by electromagnetic radiation. Electromagnetic radiation could affect the state of the captured pathogenic microorganisms and thus trigger extracellular activity of thereof. In a possible embodiment, the method includes step D) of analysing the by the captured pathogenic microorganisms extracellularly secreted and / or excreted stress responsive biomolecules. As indicated above, analysing the presence of the extracellularly secreted and / or excreted stress responsive biomolecules is an effective way to determine the presence of any pathogenic microorganisms. Subsequently, the type of pathogenic microorganisms could be analysed and / or determined. Any convention analysis method could be applied, for example a biochemical analysis methods. In case the analysis step is included, the method according to the invention could thus also be referred to as a method for analysing the presence of pathogenic microorganisms in a sample, in particular a body fluid sample.

[0028] The analysis step or step D), if applied is preferably performed by means of Raman Spectroscopy, Surface-Enhanced Raman Spectroscopy (SERS) and / or biochemical analysis. In this way, an effective and reliable analysis could be performed. It is imaginable that the prepared catch filter can be directly used as substrate in the analysis step. It is also possible that a further processing step is needed to prepare the catch filter for further analysis.

[0029] Preferably, the method comprises a step of identifying at least one type of captured pathogenic microorganisms, in particular based on the extracellularly secreted stress responsive biomolecules. This step is preferably performed after or simultaneously with step D). This identification step may for example be performed by means of Raman Spectroscopy, SERS and / or biochemical analysis. In particular, the composition of stress responsive biomolecules is characteristic for each type of pathogenic microorganism. Hence, the secreted and / or excreted extracellularly secreted stress responsive biomolecules provide information on the type, species, and / or identity of the pathogenic microorganism. Optionally, the step of identifying at least one type of captured pathogenic microorganisms is performed by comparing the identified extracellularly secreted and / or excreted stress responsive biomolecules with at least one database comprising data of the composition of stress responsive biomolecules per type of pathogenic microorganism. Possibly, at least part of at least one catch filter and / or at least part of the captured pathogenic microorganisms is provided in and / or exposed to and / or dissolved in at least one fluid. At least part of the at least one catch filter could for example be immersed in at least one fluid, in particular such that at least part of the captured pathogenic microorganisms get into contact with said fluid. As indicated above, the fluid may act as (chemical) stress stimulus. The method may comprise step E) of concentrating at least part of the at least one fluid such that the concentration of captured pathogenic microorganisms and / or the concentration of extracellularly secreted stress responsive biomolecules is increased. Step E) could for example be performed after step C). Including this step could positively affect the further analysis of the pathogenic microorganisms and / or extracellularly secreted stress responsive biomolecules. Step E) may be performed by evaporating at least a part of the at least one fluid. Step E) may be performed by heating at least part of the fluid, in particular such that at least part of the fluid evaporates. It is also possible that at least part of the catch filter is heated. This may also cause evaporation of at least part of the fluid. Hence, the method could optionally comprise a concentrating step. This is for example conceivable in case at least part of the pathogenic microorganisms and / or the secreted stress responsive biomolecules are provided and / or at least partially dissolved in a fluid. Optionally, the fluid, preferably the fluid which acts as (chemical) stress stimulus, comprises glycerol. Glycerol evaporates at higher temperatures than a majority of other components of the fluid, such as water. Hence, in case the fluid comprises glycerol the amount of fluid that evaporates during step E) can be controlled. Therewith, a well-defined volume after evaporation is obtainable. Therewith, it is provided that the catch filter does not dry out. In an alternative embodiment, at least one concentration step could be applied via freeze drying.

[0030] The method may further comprise a step F) of purifying the at least one body fluid sample for removing at least a fraction of particles from the body fluid sample. Step F) may be performed by filtering the at least one body fluid sample through at least one coarse filter. Step F) is thereby preferably performed prior to step B). Such coarse filter may be configured for removing at least a fraction of particles from the body fluid sample, in particular particles which are larger than pathogenic microorganisms. Optionally, the body fluid sample is filtered or flushed through at least one coarse filter prior to the sample being filtered through the catch filter. At least one coarse filter, if applied, is in particular configured to remove at least a fraction of particles from the sample, in particular particles which are larger than at least one analyte. This is beneficial as the analytes, in particular the pathogenic microorganisms, could be isolated and / or captured more efficiently. Within the context of this invention, when it is referred to filtering, flushing fluid through a filter and / or drawing fluid up though a filter can be meant. Within the context of this invention, filtering could also be explained as depositing of a fluid onto at least one filter. It is for example imaginable that at least part of the fluid which is to be filtered is absorbed by at least one filter.

[0031] It is conceivable that at least one coarse filter, if applied, comprises at least one coarse-grained fabric material, such as but not limited to: glass wool, cotton, synthetic wool and the like. In a beneficial embodiment, at least one coarse filter, if applied, comprises glass fiber and / or glass wool. It is for example possible that at least one coarse filter is formed by glass wool comprising or being formed by glass fiber. Glass wool and glass fiber were found to be efficient in a filtering step wherein relatively large particles are to be separated from a sample, in particular a body fluid sample. The use of glass wool and / or glass is also beneficial from economical point of view since said products are relatively cheap. It is also conceivable that at least one coarse filter comprises cellulose. Other non-limitative embodiments include at least one coarse filter comprising polycarbonate and / or a coarse filter which is at least partially made of a filter paper, coffee filter or coffee paper. It is also conceivable that at least one coarse filter as applied in a method according to the present invention comprising multiple filtering materials. Hence, the coarse filter could for example be a combination of glass wool and cellulose. It is further imaginable that at least one coarse filter comprises glass fiber and / or wherein at least one catch filter comprises a glass fiber mesh, glass wool, nylon, polyamide, cellulose acetate, polysulphone, teflon and / or cellulose.

[0032] At least one coarse filter is configured to capture particles with a particle size larger than 10 pm, preferably larger than 5 pm, more preferably larger than 2 pm, even more preferably larger than 1 pm. It is also conceivable that at least one coarse filter is configured to capture particles with a particle size (significantly) larger than 10 pm. At least on coarse filter is in particular configured to let through, particles with a particle size smaller than 5 pm, preferably smaller than 2 pm, more preferably smaller than 1 pm. In this manner, any pathogens present in the sample, which are typically smaller than said particle size, will be let through towards the catch filter step while larger contaminants will be removed from the sample. When it is referred to particle size, for example an average particle size can be meant. It is also conceivable that one coarse filter is configured to capture particles with a particle size in the range of 1 pm to 10 pm, preferably 2 pm to 5 pm. It is also conceivable that at least one coarse filter is configured to capture particles with an average width and / or length in the range of 1 pm to 10 pm.

[0033] At least one catch filter comprises glass in a preferred embodiment. It is also conceivable that at least one catch filter is at least partially made of glass. The catch filter could for example comprise glass particles, for example glass nanoparticles. At least one catch filter could for example comprise a glass (fiber) mesh. The catch filter is preferably a relatively dense filter. It is further imaginable that at least one catch filter comprises glass fiber and / or wherein at least one catch filter comprises a glass fiber mesh, glass wool, nylon, polyamide, cellulose acetate, polysulphone, polyethersulphone, teflon, polyvinylidene fluoride (PVDF), polyester (PET), polycarbonate track-etched membrane, polycarbonate (PC), and / or cellulose. It is imaginable that at least one filter is substantially hydrophobic. Preferably, at least part of at least one catch filter is substantially hydrophobic.

[0034] At least one catch filter is preferably configured to capture particles with a particle size smaller than 5 pm, preferably smaller than 3 pm, more preferably smaller than 2.5 pm, even more preferably smaller than 2 pm and most preferably smaller than 0.5 pm. In yet a further preferred embodiment, at least one catch filter is configured to capture particles with a particle size smaller than 1 pm, more in particular smaller than 0.7 pm, more in particular smaller than 0.4 pm. It may also be said that the catch filter is configured to catch particles with a predetermined particle size, for example in any of the abovementioned ranges and / or any of the ranges mentioned for the (target) analyte.

[0035] In a preferred embodiment, at least one coarse filter, if applied, and / or at least one catch filter can be present in a syringe filter. It is for example also imaginable that at least one coarse filter and / or at least one catch filter are comprised in a syringe filter. At least one coarse filter and / or at least one catch filter could for example also be a syringe filter. The filtering step could for example be applied by pushing the sample across at least one filter by the syringe. The use of at least one syringe filter in combination with a syringe could enable manual application of the method in a relatively simple manner. It is also conceivable that at least one coarse filter and / or at least one catch filter is present in a syringe filter device. Preferably, at least one filter is provided with a housing. The use of a housing could further contribute to the ease of the use of the filter. Optionally or alternatively, a filter applied within the scope of this invention could be a drip filter.

[0036] In a further possible embodiment, at least a part of the catch filter can be coated with nanoparticles, in particular SERS-active nanoparticles to form a SERS substrate. The nanoparticles enable that the catch filter can be used in further analysis steps, in particular SERS analysis. This results in enhanced possibilities to analyse the presence of any pathogenic microorganisms. At least part of the nanoparticles could be metal nanoparticles. It is also conceivable that substantially all nanoparticles are metal nanoparticles. Preferably at least part of the nanoparticles are noble metal nanoparticles. Metal nanoparticles, and in particular noble metal nanoparticles were found to be rather efficient for use in Raman / SERS spectroscopy. It is for example conceivable that at least part of the nanoparticles is selected from the group of silver, nickel, aluminium, gold, platinum, palladium, titanium, copper, cobalt, zinc, and / or combinations thereof. It is for example also conceivable that at least part of the nanoparticles comprises an alloy, in particular of any of the listed preferred metals. In a preferred embodiment, at least part of the nanoparticles are gold nanoparticles. It is also conceivable that at least part of the nanoparticles comprises a combination of (noble) metal and silica, such as silica shelled (metal) nanoparticles. It is also imaginable that at least part of the nanoparticles are metal nanoparticles comprising a porous silica shell. In a possible embodiment, the method comprises step G) of filtering at least one solution comprising nanoparticles, in particular SERS-active nanoparticles, through the at least one catch filter such that at least part of the catch filter will be coated with nanoparticles, in particular SERS-active nanoparticles. The at least partially coated catch filter could subsequently be used as SERS substrate. Optionally, the at least one solution comprising nanoparticles is configured for applying at least one stimulus, in particular at least one (chemical) stress stimulus, when applied to at least a part of the at least one catch filter and / or to the fraction of pathogenic microorganisms, if captured. As indicated above, the solution may act as (chemical) stress stimulus. If the method comprises step E), at least part of the solution is concentrated such that the concentration of captured pathogenic microorganisms and / or the concentration of extracellularly secreted stress responsive biomolecules is increased.

[0037] The invention also relates to a device for processing at least one body fluid sample for pathogenic microorganism analysis, comprising at least one housing, comprising at least one retaining structure for retaining at least one catch filter, and at least one fluid reservoir for receiving at least one fluid, optionally, at least one catch filter configured to capture at least a fraction of pathogenic microorganisms from a body fluid sample, wherein at least one housing is a modular housing which is adjustable at least between a filtering configuration wherein at least one body fluid can be filtered through at least one catch filter retained in the at least one retaining structure and a processing configuration wherein at least one retaining structure is positioned substantially adjacent to at least part of the at least one fluid reservoir such that fluid received in the at least one fluid reservoir can get into contact with at least part of at least one catch filter retained in the at least one retaining structure.

[0038] The device according to the present invention can be configured to perform a method according to the present invention. The method according to the present invention could be performed by making use of a device according to the present invention. The device in particular enables that processing at least one body fluid sample for pathogenic microorganism analysis can done in a relatively simple and effective manner. Alternatively, the device could also be used for other applications in fluid analysis, for example the analysis of at least one analyte in a fluid.

[0039] The use of a modular housing which is adjustable between a filtering configuration and a processing configuration enables that the at least one catch filter can be effectively used in the processing of the body fluid sample and that subsequently preparation of the captured pathogenic microorganism analysis can be achieved such that analysis thereof can be done. The device according to the present invention can be adjusted to a processing configuration wherein at least part of at least one retaining structure is positioned substantially adjacent to at least part of the at least one fluid reservoir in particular such that fluid received in the at least one fluid reservoir can get into contact with at least part of at least one catch filter retained in the at least one retaining structure and / or with at least part of the captured pathogenic microorganism. It is for example possible that at least part of at least one retaining structure is positioned substantially parallel to at least part of the at least one fluid reservoir.

[0040] It is beneficial if at least one retaining structure and at least one fluid reservoir are mutually displaceable. It is also imaginable that at least one retaining structure is displaceable at least between the filtering configuration and the processing configuration. At least one fluid reservoir could be substantially stationary received in the housing. In yet another possible embodiment, it is also imaginable that I at least one retaining structure and at least one fluid reservoir are displaceable. Alternatively, at least one fluid reservoir could be displaceable and at least one retaining structure could be substantially stationary positioned within the housing. It is for example imaginable that at least one retaining structure and / or at least one fluid reservoir is slidable between the filtering configuration and the processing configuration.

[0041] The device could optionally comprise at least one catch filter configured to capture at least a fraction of pathogenic microorganisms from a body fluid sample. Any of the possible catch filters as described for the corresponding method according to the present invention could be applied. The at least one retaining structure is in particular configured for retaining at least one catch filter. It is for example possible that at least one retaining structure could clampingly engage at least part of at least one catch filter.

[0042] At least fluid reservoir is in particular configured for receiving at least one fluid and / or at least one (chemical) substance. It is for example possible that at least one fluid reservoir is configured to receive at least 5 ml fluid, in particular at least 10 ml more in particular at least 20 ml. It is also conceivable that at least one fluid reservoir is configured to receive at most 100 ml fluid, in particular at most 50 ml, more in particular at most 25 ml.

[0043] It is possible that, at least in the processing configuration, at least part of at least one catch filter retained in the retaining structure at least partially covers the fluid reservoir. The fluid reservoir could for example define at least one fluid opening. It is for example possible that at least part of at least one catch filter retained in the retaining structure at least partially, and preferably fully covers, at least one fluid opening of the fluid reservoir at least in the processing configuration. Hence, the housing is preferably configured such that at least part of at least one catch filter retained in the retaining structure at least partially, and preferably fully covers, at least one fluid opening of the fluid reservoir at least in the processing configuration. It is imaginable that at least one catch filter retained in the retaining structure is positioned at a distance from at least one fluid reservoir and in particular at least one opening of at least one fluid reservoir at least in the filtering configuration.

[0044] The device could also comprise at least one sealing member configured to seal at least part of the fluid reservoir in particular at least one opening of the fluid reservoir. It is also possible that the device, and in particular the housing, and more in particular at least one fluid reservoir and / or at least one retaining structure, comprises at least one sealing member configured to couple the fluid reservoir and the retaining structure in a liquid-proof or liquid-tight manner, preferably at least in the processing configuration. It is for example possible that at least part of at least one sealing member surrounds or encloses at least part of the catch filter retained in the retaining structure. It is possible that at least one sealing member is a releasable sealing member. It is also conceivable that at least one sealing member forms integral part of the housing, in particular of the retaining structure and / or the fluid reservoir.

[0045] It is imaginable that at least part of the fluid reservoir is detachably with respect to the housing. It is also possible that at least part of retaining structure is detachable with respect to the housing. For example, the fluid reservoir and / or the retaining structure can be detachably connected to the housing. At least part of the fluid reservoir and / or the retaining structure being detachable is beneficial for cleaning purposes. In case the device is used multiple times, the device need to be fully cleaned prior to a new cycle of use. During use, for example when a method according to the invention is applied, it is highly desired that the device is not contaminated as this could negatively affect the analysis of any pathogenic microorganisms.

[0046] In a possible embodiment, at least part of the at least one fluid reservoir comprises at least one substantially translucent and / or substantially transparent portion. It is beneficial if the fluid reservoir comprises at least one translucent and / or transparent portion for enabling visual inspection and / or analysis purposes. It is possible that the presence of pathogenic microorganisms is analysed via fluid present in the fluid reservoir. It is in particular interesting if a portion of the fluid reservoir located at an opposite side of at least a part of the retaining structure, more particular located at an opposite side of the catch filter if retained in the retaining structure, is at least partially translucent and / or transparent. It is for example possible that at least one fluid reservoir comprises a substantially transparent and / or substantially translucent side wall. It is also possible that the fluid reservoir is formed of a substantially transparent and / or substantially translucent material.

[0047] The housing may comprise at least one application opening. At least one application opening is preferably at least partially aligned with a catch filter retained in the retaining structure at least one in the filtering configuration. At least one application opening can be configured for passaging at least one body fluid sample trough or to the catch filter in the filtering configuration. At least one application opening could also be referred to as a syringe opening. The at least one syringe opening could be configured for receiving at least part of at least one syringe. It is configured that at least one syringe could be connected to at least one housing in particular via at least one application opening. It is also possible that the device comprises at least one sample applicator for applying at least one body fluid sample to the catch filter at least in the filtering configuration. At least one sample applicator is connected or connectable to at least one application opening.

[0048] In a possible embodiment, the housing defines at least one accommodation space configured to at least partially accommodate at least one coarse filter. At least one coarse filter is preferably configured for removing at least a fraction of particles from at least one body fluid sample, in particular particles which are larger than pathogenic microorganisms. In a further preferred embodiment, at least one accommodation space is at least partially aligned with the retaining structure configured to retain at least one catch filter, at least in the filtering configuration. It is also imaginable that the device, and in particular the housing thereof, comprises at least one secondary retaining structure configured to retain at least part of at least one coarse filter. It is imaginable that the secondary retaining structure is positioned adjacent to, for example substantially parallel to, the (primary) retaining structure such that at least one coarse filter retained in the secondary retaining structure is substantially parallel to at least one catch filter retained in the (primary) retaining structure. The coarse filter and the catch filter are preferably at least partially aligned during use. It is imaginable that the coarse filter is provided at an upstream side from the catch filter in respect of an application direction of the body fluid sample to the catch filter. The body fluid sample is preferably first filtered through the coarse filter and thereafter through the catch filter. The body fluid can in fact be filtered substantially simultaneously through at least one coarse filter and at least one catch filter. In a possible embodiment, it is also imaginable that the retaining structure configured to retain at least one catch filter and at least one coarse filter.

[0049] Thee device, and in particular the housing may comprise multiple openings. It is for example possible that the housing comprises at least one processing opening. Such processing opening is preferably at least partially aligned with the fluid reservoir in the processing configuration for passaging fluid to the fluid reservoir in the processing configuration. The device may further comprise at least one fluid applicator for applying fluid to the at least one fluid reservoir in the processing configuration. The device can also be configured for co-action with at least one fluid applicator, which is in particular configured for applying fluid to the at least one fluid reservoir in the processing configuration. In a possible embodiment, at least one fluid applicator comprises a fluid configured to apply a stress stimulus to the at least part of at least one catch filter retained in the at least one retaining structure, in particular to the captured fraction of pathogenic microorganisms from a body fluid sample if present in the body fluid sample. At least one fluid applicator may also comprise a solution comprising SERS-active nanoparticles to be filtered through the catch filter retained in the retaining structure such that at least part of the catch filter will be coated with SERS-active nanoparticles thereby forming a SERS substrate. At least one fluid applicator can be connected or connectable to at least one processing opening for passaging fluid from the fluid applicator to the fluid reservoir in the processing configuration. In a possible embodiment, the retaining structure comprises at least one inlet which is at least partially aligned with and / or joined to the fluid reservoir in the processing configuration, in particular for passaging fluid to the fluid reservoir in the processing configuration. It is conceivable that the processing opening and the inlet together form a fluid conduct in the processing configuration for guiding fluid to and / or from the fluid reservoir.

[0050] The device may further comprise at least one absorbing body for absorbing a fraction of fluid. This is in particular useful when the fluid is present in excess. At least one absorbing body may comprise at least one fluid absorbing material. This can for example be a sponge or a sponge-like material. It is preferred that at least one absorbing body is provided adjacent to at least part of the fluid reservoir and / or adjacent to at least one (fluid) inlet. At least one absorbing body is for example provided at an end side of the housing.

[0051] The invention also relates to a device for processing at least one body fluid sample for pathogenic microorganism analysis, comprising at least one housing, comprising at least one retaining structure for retaining at least one catch filter, and at least one stress applicator for applying at least one stress stimulus to at least a part of at least one catch filter retained in at least one retaining structure, optionally, at least one catch filter configured to capture at least a fraction of pathogenic microorganisms from a body fluid sample, wherein at least one housing is a modular housing which is adjustable between a filtering configuration wherein at least one body fluid can be filtered through at least one catch filter retained in the at least one retaining structure and a processing configuration wherein at least one retaining structure is positioned such that at least part of at least one stress applicator is, directly or indirectly, in contact with at least part of at least one retaining structure and / or at least one catch filter retained in the at least one retaining structure.

[0052] At least one stress applicator can for example be configured to apply at least one chemical, mechanical and / or electrical stress stimulus. Any of the above described embodiments of the method and device according to the invention can be combined with this embodiment. Similar features of this device and the above described embodiments of the method and device have similar advantages.

[0053] The invention further relates to the use of (at least part of) a device according to the present invention, in particular in a Raman Spectrometer for analysing extracellularly secreted stress responsive biomolecules of pathogenic microorganisms.

[0054] The invention will be further elucidated by means of the following non-limitative clauses.

[0055] 1 . Method for processing at least one body fluid sample for pathogenic microorganism analysis, comprising the steps of:

[0056] A) providing at least one body fluid sample,

[0057] B) filtering the at least one body fluid sample through at least one catch filter which is configured for capturing at least a fraction of pathogenic microorganisms from the at least one body fluid sample if present in the body fluid sample, and

[0058] C) applying at least one stress stimulus to at least part of the at least one catch filter, in particular to the fraction of pathogenic microorganisms, such that at least a fraction of the captured pathogenic microorganisms extracellularly secrete stress responsive biomolecules.

[0059] 2. Method according to clause 1 , wherein the steps A) to C) are subsequent steps.

[0060] 3. Method according to any of the preceding clauses, wherein during step C) at least one chemical stress stimulus is applied to at least part of the at least one catch filter.

[0061] 4. Method according to clause 3, wherein the chemical stress stimulus comprises at least one fluid and / or at least one chemical substance, preferably chosen from the group consisting of: water, demi-water, an acid, a base, an antimicrobial substance and / or a combination thereof. 5. Method according to any of the preceding clauses, wherein during step C) at least one mechanical stress stimulus is applied to at least part of the at least one catch filter.

[0062] 6. Method according to clause 5, wherein the mechanical stress stimulus is a mechanical force.

[0063] 7. Method according to any of the preceding clauses, wherein during step C) at least one electrical stress stimulus is applied to at least part of the at least one catch filter.

[0064] 8. Method according to any of the preceding clauses, wherein during step C) at least one stress stimulus is applied by a change in temperature and / or by electromagnetic radiation.

[0065] 9. Method according to any of the preceding clauses, comprising a step D) of analysing the by the captured pathogenic microorganisms extracellularly secreted stress responsive biomolecules.

[0066] 10. Method according to clause 9, wherein step D) is performed by means of Raman Spectroscopy, Surface-Enhanced Raman Spectroscopy (SERS) and / or biochemical analysis.

[0067] 11 . Method according to any of the preceding clauses, comprising a step of identifying at least one type of the captured pathogenic microorganism, in particular based on the extracellularly secreted stress responsive biomolecules.

[0068] 12. Method according to any of the preceding clauses, wherein at least part of the catch filter and / or at least part of the captured pathogenic microorganisms is provided in and / or dissolved in at least one fluid.

[0069] 13. Method according to clause 12, comprising step E) of concentrating at least part of the at least one fluid such that the concentration of captured pathogenic microorganisms and / or the concentration of extracellularly secreted stress responsive biomolecules is increased. 14. Method according to clause 13, wherein step E) is performed by heating.

[0070] 15. Method according to any of the preceding clauses, comprising a step F) of filtering the at least one body fluid sample through at least one coarse filter which is configured for removing at least a fraction of particles from the body fluid sample, in particular particles which are larger than pathogenic microorganisms.

[0071] 16. Method according to clause 15, wherein step F) is performed prior to step B).

[0072] 17. Method according to any of the preceding clauses, wherein at least a part of the catch filter is coated with nanoparticles, in particular SERS-active nanoparticles to form a SERS substrate.

[0073] 18. Method according to any of the preceding clauses, comprising a step G) of filtering at least one solution comprising nanoparticles, in particular SERS-active nanoparticles through the catch filter such that at least part of the catch filter will be coated with said nanoparticles.

[0074] 19. Device for processing at least one body fluid sample for pathogenic microorganism analysis, comprising:

[0075] - at least one housing, comprising: o at least one retaining structure for retaining at least one catch filter, and o at least one fluid reservoir for receiving at least one fluid,

[0076] - optionally, at least one catch filter configured to capture at least a fraction of pathogenic microorganisms from a body fluid sample, wherein at least one housing is a modular housing which is adjustable between:

[0077] - a filtering configuration wherein at least one body fluid can be filtered through at least one catch filter retained in the at least one retaining structure; and

[0078] - a processing configuration wherein at least one retaining structure is positioned adjacent to at least part of the at least one fluid reservoir such that fluid received in the at least one fluid reservoir can get into contact with at least part of at least one catch filter retained in the at least one retaining structure.

[0079] 20. Device according to clause 19, wherein at least one retaining structure is displaceable at least between the filtering configuration and the processing configuration.

[0080] 21 . Device according to any of the clauses 19 to 20, wherein in the processing configuration at least one catch filter retained in the retaining structure at least partially covers the fluid reservoir.

[0081] 22. Device according to any of the clauses 19 to 21 , wherein comprising at least one sealing member configured to couple the fluid reservoir and the retaining structure in a liquid-proof or liquid-tight manner, preferably at least in the processing configuration.

[0082] 23. Device according to any of the clauses 19 to 22, wherein the fluid reservoir and / or the retaining structure are detachably connected to the housing.

[0083] 24. Device according to any of the clauses 19 to 23, wherein at least part of the at least one fluid reservoir comprises a translucent or transparent portion.

[0084] 25. Device according to any of the clauses 19 to 24, wherein the housing comprises at least one application opening which is at least partially aligned with the catch filter retained in the retaining structure in the filtering configuration for passaging at least one body fluid sample to the catch filter in the filtering configuration.

[0085] 26. Device according to any of the clauses 19 to 25, comprising at least one sample applicator for applying at least one body fluid sample to the catch filter at least in the filtering configuration.

[0086] 27. Device according to clause 25 and clause 26, wherein the sample applicator is connected or connectable to at least one application opening. 28. Device according to any of the clauses 19 to 27, wherein the housing defines at least one accommodation space configured to at least partially accommodate at least one coarse filter which is configured for removing at least a fraction of particles from at least one body fluid sample, in particular particles which are larger than pathogenic microorganisms, and preferably wherein the at least one accommodation space is at least partially aligned with the retaining structure configured to retain at least one catch filter, at least in the filtering configuration.

[0087] 29. Device according to any of the clauses 19 to 28, wherein the housing comprises at least one processing opening which is at least partially aligned with the fluid reservoir in the processing configuration for passaging fluid to the fluid reservoir in the processing configuration.

[0088] 30. Device according to any of the clauses 19 to 29, comprising at least one fluid applicator for applying fluid to the at least one fluid reservoir in the processing configuration.

[0089] 31 . Device according to clause 30, wherein at least one fluid applicator comprises a fluid configured to apply a stress stimulus to the at least part of at least one catch filter retained in the at least one retaining structure, in particular to the captured fraction of pathogenic microorganisms from a body fluid sample if present in the body fluid sample.

[0090] 32. Device according to clause 30 or clause 31 , wherein at least one fluid applicator comprises a solution comprising SERS-active nanoparticles to be filtered through the catch filter retained in the retaining structure such that at least part of the catch filter will be coated with SERS-active nanoparticles thereby forming a SERS substrate.

[0091] 33. Device according to clause 29 and any of the clauses 30 to 32, wherein at least one fluid applicator is connectable to at least one processing opening for passaging fluid from the fluid applicator to the fluid reservoir in the processing configuration. 34. Device according to any of the clauses 19 to 33, wherein the retaining structure comprises at least one inlet which is at least partially aligned with the fluid reservoir in the processing configuration for passaging fluid to the fluid reservoir in the processing configuration.

[0092] 35. Device according to clause 29 and clause 34, wherein the processing opening and the inlet together form a fluid conduct in the processing configuration for guiding fluid to the fluid reservoir.

[0093] 36. Device according to any of the clauses 19 to 35, comprising at least one absorbing body for absorbing a fraction of fluid.

[0094] 37. Device according to clause 36, wherein at least absorbing body is provided adjacent to the fluid reservoir.

[0095] 38. Use of the sample processing device according to any of the clauses 19 to 37 in a Raman Spectrometer for analysing extracellularly secreted stress responsive biomolecules of pathogenic microorganisms.

[0096] The invention will be further elucidated by several examples and with reference to the appended figures, wherein:

[0097] - Figure 1 schematically shows a perspective view of a device for processing at least one body fluid sample for pathogenic microorganism analysis according to the present invention,

[0098] - Figure 2 schematically shows a cross-sectional view of the device of figure 1 in the filtering configuration,

[0099] - Figures 3a and 3b schematically show a cross-sectional view of the device of figure 1 in the processing configuration,

[0100] - Figures 4a-4d schematically show another embodiment of a device for processing at least one body fluid sample for pathogenic microorganism analysis according to the invention, and

[0101] - Figure 5 schematically shows a method for processing at least one body fluid sample for pathogenic microorganism analysis according to the invention. Within these figures, similar reference numbers correspond to similar or equivalent elements or features.

[0102] Figure 1 schematically shows a perspective view of a device 1 for processing at least one fluid sample S, in particular a body fluid sample S, for analyte analysis, in particular pathogenic microorganism analysis, according to the present invention. The device 1 comprises a housing 2 which comprises a retaining structure 3 for retaining a catch filter (shown in figure 2). In the shown embodiment, a sample applicator 6 is connected to the housing 2 of the device 1 . The shown sample applicator 6 is in the shown embodiment a syringe, which comprises a receiving section 6a for (filtered) body fluid sample S and an aspiration tube 6b an aspiration tube 6b for passaging body fluid sample S to, and through, the housing 2. It is possible that only the receiving section 6a or the aspiration tube 6b is connected to the housing 2. In another embodiment, the receiving section 6a may be configured to apply a body fluid sample S to the housing 2, in particular to the catch filter. Optionally, the aspiration tube 6b is configured to discharge filtered body fluid sample S from the housing, in particular from the catch filter. The syringe 6a and the aspiration tube 6b are at least partially aligned, in particular such that fluid drawn up from the aspiration tube 6b can flow directly towards the syringe 6a. The syringe 6a and the aspiration tube 6b are provided on opposing sides of the housing 2. The housing 2 and / or the sample applicator 6 may further comprise a coarse filter 7 for removing at least a fraction of particles from the body fluid sample S, in particular particles which are larger than pathogenic microorganisms.

[0103] Optionally, the coarse filter 7 is connected to the aspiration tube 6b.

[0104] The shown device 1 further comprises a fluid applicator 8 for applying fluid F to a fluid reservoir (shown in figure 2). The shown fluid applicator 8 is connected to the housing 2. In the shown embodiment, the fluid applicator 8 is connected to the housing 2 at a distance from the sample applicator 6. The shown fluid applicator 8 comprises at least one fluid retainer 8a’, 8a” configured for retaining at least one fluid F to be applied to the housing 2, in particular to the fluid reservoir. In the shown embodiment, the fluid retainer 8a’, 8a” is a syringe. The shown fluid applicator 8 comprises two fluid retainers 8a’, 8a” wherein a first fluid retainer 8a’ is configured to apply at least one fluid to the housing 2, in particular to the fluid reservoir, and a second fluid retainer 8a” is configured to receive applied fluid F from the housing 2, in particular from the fluid reservoir. It is imaginable that the second fluid container 8a” comprises an aspiration or suction mechanism configured to actively aspirate fluid F from the housing 2, in particular form the fluid reservoir, to the fluid retainer 8a”. The fluid applicator 8 of the shown embodiment further comprises a fluid applicator holder 8b for holding the fluid applicator 8 on the housing 2. The fluid applicator holder 8b may be configured to apply a force on a plunger of the first fluid retainer 8a’. Optionally, the fluid applicator holder 8b is configured to apply a predetermined force to the plunger of the first fluid retainer 8a’ to control the amount of fluid F to be applied to the housing 2, in particular to the fluid reservoir. The fluid applicator holder 8b of the shown embodiment is in contact with part of a covering element 14 to limit movement of the covering element 14, in particular in a longitudinal direction of the housing 2. The device 1 , and in particular the housing 2, furthermore comprises an absorbing body 10 configured to absorb a fraction of fluid F and / or body fluid sample S, in particular when present in excess. The absorbing body 10 is preferably provided at an end side of the housing 2. In the shown embodiment, the absorbing body 10 is provided next to or near the fluid applicator 8. The absorbing body 10 may for example comprise a sponge.

[0105] Figure 2 schematically shows a cross-sectional view of the device 1 according to figure 1 in the filtering configuration. The device 1 comprises a housing 2 which comprises a retaining structure 3 for retaining a catch filter 4. The catch filter 4 is configured to capture at least a fraction of pathogenic microorganisms from the body fluid sample S if present. In the shown embodiment, the body fluid sample S is filtered through the catch filter 4 retained in the retaining structure 3. A portion of the shown retaining structure 3 and / or the housing 2 at the location of the filtering configuration comprises a first sealing member 17 to liquid-tight or liquid-proof enclose the catch filter 4 retained in the retaining structure 3. Therewith, leakage of body fluid sample S into the remaining of the housing 2 is limited or prevented. The body fluid sample S may be applied by means of a sample applicator 6. In the shown embodiment, the sample applicator 6 is connected to the housing 2. The housing 2 may comprise an application opening 11 for passaging a body fluid sample S. The application opening 11 is preferably at least partially aligned with the catch filter 4 in the filtering configuration, in particular such that the body fluid sample S can be filtered through the catch filter 4. In the shown embodiment, the housing 2 comprises two application openings 11 . The application openings 11 may together form a fluid conduct for guiding the body fluid sample S into and away from the housing 2, in particular such that the body fluid sample S is filtered through the catch filter 4. In the filtering configuration, the application openings 11 are preferably at least partially aligned with at least a part of the catch filter 4. In the filtering configuration, the sample applicator 6 is preferably at least partially aligned with the catch filter 4, such that the body fluid sample S can be filtered through the catch filter 4. The sample applicator 6 may be an aspiration device, such as a syringe. The shown sample applicator 6 comprises a receiving section 6a for receiving the filtrate of the filtered body fluid sample S and an aspiration tube 6b for passaging body fluid sample S to the housing 2, in particular to the catch filter 4. In the shown embodiment, the aspiration tube 6b is connected to the housing 2 at an opposite side of the receiving section 6a the aspiration tube 6b. The receiving section 6a and the aspiration tube 6b are connected to the housing 2 via application openings 11 . The housing 2 and / or the sample applicator 6 may further comprise a coarse filter 7 for removing at least a fraction of particles from the body fluid sample S, in particular particles which are larger than pathogenic microorganisms. The coarse filter 7 and the catch filter 4 are preferably at least partially aligned. It is imaginable that the coarse filter 7 is provided at an upstream side from the catch filter 4 in respect of an application direction of the body fluid sample S to the catch filter 4. Optionally, the catch filter 4 can be provided at a downstream side of the coarse filter 7 in respect of an application direction of the body fluid sample S to the catch filter 4. The body fluid sample S is preferably first filtered through the coarse filter 7 and thereafter through the catch filter 4. In the shown embodiment, at least a part of the coarse filter 7 is connected to an application opening 11 of the housing 2. In the shown embodiment, the housing 2 comprises an accommodation space 12 for accommodating a coarse filter 7. In particular at least one side wall 13 of the accommodation space 12 comprises an application opening 11. More in particular, two opposite side walls 13 of the accommodation space 12 comprise an application opening 11. The application openings 11 at the opposite side walls 13 of the accommodation space 12 are at least partially aligned, such that body fluid sample S can be passed through the coarse filter 7 in the accommodation space 12.

[0106] The housing 2 further comprises a fluid reservoir 5 for receiving at least one fluid F.

[0107] The housing 2 is adjustable from the shown filtering configuration to a processing configuration, as shown in more detail in figures 3a and 3b. The retaining structure 3 is moveable, in particular slideable, at least between the filtering configuration and the processing configuration. The fluid reservoir 5 is in the shown embodiment enclosed by the retaining structure 3 and a covering element 14. The covering element 14 may comprise a transparent or translucent cover 15. The covering element 14 and / or at least a part of the housing 2 and / or at least a part of the retaining structure 3 comprise a second sealing member 16 configured to liquidproof or liquid-tight enclose the fluid reservoir 5. The shown retaining structure 3 comprises an inlet 9a for passaging fluid F, in particular when the device 1 is positioned in the processing configuration (see figures 3a and 3b). The shown inlet 9a is provided near, or substantially adjacent to, the catch filter 4 if present in the processing configuration. The housing 2 may comprise a processing opening 9b for passaging fluid F, in particular when the device 1 is positioned in the processing configuration. The processing opening 9b may at least partially be fluidly connected to the fluid reservoir 5. The inlet 9a of the retaining structure 3 and the processing opening 9b of the housing 2 are at least partially aligned in the processing configuration for allowing fluid F to passage to the fluid reservoir 5 in the processing configuration. In the shown embodiment, the inlet 9a and the processing opening 9b form a fluid conduct for passaging fluid F to the fluid reservoir 5. In the shown embodiment, a fluid applicator 8 comprising fluid F is connected to the passage opening 9b of the housing 2.

[0108] Figures 3a and 3b schematically show a cross-sectional view of the device 1 of figures 1 and 2 in the processing configuration. In the processing configuration, the retaining structure 3 is positioned substantially parallel to at least part of the fluid reservoir 5 such that fluid F received in the fluid reservoir 5 can get into contact with at least part of at least one catch filter 4 retained in the at least one retaining structure 3. Compared to the filtering configuration from figure 2, the catch filter 4 and the housing 2 are mutually displaced, in particular in a longitudinal direction of the housing 2. The catch filter 4 and the covering element 14 mutually enclose the fluid reservoir 5. In the shown figures, the processing opening 9b and the inlet 9a are aligned. Therewith, the processing opening 9b and the inlet 9a form a fluid conduct for passaging fluid F from the fluid applicator 8 to the fluid reservoir 5.

[0109] Figure 3a shows the processing configuration prior to the application of fluid F to the fluid reservoir 5. As can be observed, the fluid applicator 8 is connected to the processing opening 9b. The first fluid retainer 8a’ comprises fluid F to be applied to the fluid reservoir 5, in particular to the catch filter 4. The retaining structure 3 and / or the covering element 14 may comprise a locking element 18 configured to lock or connect the retaining structure 3 and the covering element 14 at least in the processing configuration. In the shown embodiment, the retaining structure 3 comprises a first locking part 18a and the covering element 14 comprises a second locking part 18b, wherein the first locking part 18a and the second locking part 18b are complementary to mutually connect or lock the retaining structure 3 and the covering element 14.

[0110] Figure 3b shows the processing configuration wherein fluid F from the first fluid retainer 8a’ is applied to the fluid reservoir 5. The fluid F is pushed out of the first fluid retainer 8a’ by a first force F1 . The fluid F first passages the processing opening 9b of the housing 2. Thereafter, the fluid F passages the inlet 9a of the retaining structure 3. The second sealing member 16 encloses the fluid reservoir 5. Hence, the fluid F is kept in the fluid reservoir 5. In the shown embodiment, the fluid F is in contact with both the covering element 14 and the catch filter 4. The second fluid retainer 8a” aspirates or sucks at least a fraction of the fluid F from the fluid reservoir 5 into the second fluid retainer 8a” with a second force F2. Optionally, the first force F1 and the second F2 are substantially equal. The fluid F is at least temporarily in contact with captured pathogenic microorganisms on the catch filter 4 and / or in the fluid reservoir 5 and / or on the covering element 14, if present. Thereafter, the fluid F may be drawn through the catch filter 4 into the second fluid retainer 8a”. The fluid F preferably is or comprises a substance configured to apply a stress stimulus to the at least part of at least one catch filter 4 retained in the retaining structure 3, in particular to the captured fraction of pathogenic microorganisms from a body fluid sample S if present in the body fluid sample S. Optionally, the fluid F comprises a solution comprising SERS-active nanoparticles to be filtered through the catch filter 4 retained in the retaining structure 3 such that at least part of the catch filter 4 will be coated with SERS-active nanoparticles.

[0111] Figures 4a-4d schematically show another embodiment of a device 40 for processing at least one body fluid sample S for pathogenic microorganism analysis according to the invention. The device 40 comprises a housing 42 comprising at least one retaining structure 43 for retaining at least one catch filter 44. The housing 2 further comprises at least one stress applicator 45, 47, 48 for applying at least one stress stimulus to at least a part of at least one catch filter 44 retained in at least one retaining structure 43. In the shown embodiment, the catch filter 44 is configured to capture at least a fraction of pathogenic microorganisms from a body fluid sample S if present in the body fluid sample S. The housing 42 of the shown embodiment comprises an application opening 41 configured for applying of at least one body fluid sample S to the catch filter 44 retained in the retaining structure 44. The housing 42 further comprises a processing opening 49. In another embodiment, it is imaginable that the housing 42 comprises one of the application opening 41 and processing opening 49. The retaining structure 43 may comprise a slot 50 or a manoeuvre space 50 which divides the retaining structure 43 in two sections 43a, 43b. The slot 50 or manoeuvre space 50 allows the sections 43a, 43b to move or manoeuvre relatively to each other, in particular relative to the slot 50 or manoeuvre space 50. The slot 50 or manoeuvre space 50 extends in a longitudinal direction of the retaining structure 43. The slot 50 or manoeuvre space 50 preferably extends between the catch filter 44 and at least one lateral side of the retaining structure 43. The housing 42 is adjustable between a filtering configuration and a processing configuration. Figure 4a shows the device 40 in the filtering configuration and figures 4b-4d show possible embodiments of in the processing configuration.

[0112] Figure 4a schematically shows the housing 42, or the device 40, in the filtering configuration, wherein at least one body fluid S is filtered through the catch filter 44. Preferably, the catch filter 44 is at least partially aligned with the application opening 41. The housing 42 may be adjusted between the filtering configuration and the processing configuration (see figures 4b-4d) by mutually displacing the housing 42 and the retaining structure 43. This may for example be done by displacing or sliding the retaining structure 43 in a longitudinal direction of the housing 42. Preferably at least such that at least a part of the retaining structure 43 and / or the catch filter 44 retained in the retaining structure 43 is at least partially aligned with the processing opening 49 in the processing configuration and such that at least a part of the retaining structure 43 and / or the catch filter 44 retained in the retaining structure 43 is at least partially aligned with the application opening 41.

[0113] Figures 4b-4c schematically show the housing 42, or the device 40, in the processing configuration, wherein the retaining structure 43 is positioned such that at least part of at least one stress applicator 45, 47, 48 is in contact with at least part of at least one retaining structure 43 and / or at least one catch filter 44 retained in the at least one retaining structure 43.

[0114] Figure 4b schematically shows an embodiment wherein the stress applicator 45 is a chemical stress applicator 45 configured to apply at least one chemical stress stimulus to at least a portion of at least one catch filter 44 retained in the retaining structure 43, in particular to at least a portion of the captured pathogenic microorganisms. Therewith, the environment of the captured pathogenic microorganisms changes chemically. Preferably, the chemical stress applicator 45 comprises at least one fluid and / or a chemical substance.

[0115] Figure 4c schematically shows an embodiment wherein the stress applicator

[0116] 47 is an electrical stress applicator 47 configured to apply at least one electrical stress stimulus is applied to at least a portion of the at least one catch filter 44 retained in the retaining structure 43, in particular to at least a portion of the captured pathogenic microorganisms. The electrical stress applicator 47 may for example provide a current onto at least a portion of the catch filter 44. Optionally, the catch filter 44 is made from a conductive material configured to conduct a current applied by the electrical stress applicator 47.

[0117] Figure 4d schematically shows an embodiment wherein the stress applicator

[0118] 48 is a mechanical stress applicator 48 configured to apply at least one mechanical stress stimulus to at least a portion of the at least one catch filter 44 retained in the retaining structure 43, in particular to at least a portion of the captured pathogenic microorganisms. Therewith, the environment of the captured pathogenic microorganisms changes mechanically. In the shown embodiment, the mechanical stress stimulus is a mechanical force Fp. The retaining structure 43 of the shown embodiment comprises a slot 50 or a manoeuvre space 50 which divides the retaining structure 43 in two sections 43a, 43b. In the shown embodiment, a mechanical force Fp is applied on at least one of the retaining structure sections 43a, 43b. The shown mechanical force Fp is a pulling force Fp. By applying the pulling force Fp to at least one of the two retaining structure sections 43a, 43b, the two retaining structure sections 43a, 43b relatively move away from each other and the slot 50 becomes wider. The catch filter 44 retained in the retaining structure 43 is also exposed to a pulling force Fp. Hence, the catch filter 44 stretches in a direction of the pulling force Fp. Therewith, a mechanical stress stimulus is applied to at least a portion of the catch filter 44. In another embodiment, it is imaginable that another mechanical force Fp is applied to the catch filter 44. For example, by a pushing force wherein the retaining structure sections 43a, 43b are pushed relatively towards each other, in particular in the direction of the slot 50 or manoeuvre space 50. By applying a pushing force to at least one of the two retaining structure sections 43a, 43b, the two retaining structure sections 43a, 43b relatively move towards each other and the slot 50 becomes smaller. Hence, the catch filter 44 retained in the retaining structure 43 is also exposed to the pushing force. Hence, the catch filter 44 may compress in a direction of the pushing force.

[0119] Optionally, one or multiple different stress stimuli are applied to at least part of at least one retaining structure and / or at least one catch filter retained in the at least one retaining structure, in particular to the at least a portion of the captured pathogenic microorganisms.

[0120] Figure 5 schematically shows a method for processing at least one fluid sample S for analyte analysis according to the invention. In step A) the fluid sample S is provided. The fluid sample is preferably a body fluid sample S, such as blood, urine, sweat, salvia, and the like. In step B) the (body) fluid sample S is filtered through at least one catch filter 104. The catch filter 4 is configured to capture at least a fraction of viable cells, in particular pathogenic microorganisms 100a, 100b, such as fungi, bacteria, and the like. In step C) at least one environmental change is applied to at least a portion the captured viable cells. In the shown embodiment, a (stress) stimulus 102 is applied to at least a portion of the at least one catch filter 104, in particular to the fraction of pathogenic microorganisms 100a, 100b. The (stress) stimulus 102 may be an electrical (stress) stimulus 103, a chemical (stress) stimulus and / or a mechanical (stress) stimulus 105. Optionally, the (stress) stimulus is a change in temperature or electromagnetic radiation. The application of at least one stress stimulus may result in a change in environment of the pathogenic microorganisms 100a, 100b. As a result thereof, at least a fraction of the captured pathogenic microorganisms 100a, 100b extracellularly secrete and / or excrete stress responsive biomolecules 101. The stress responsive biomolecules 101 may be metabolites. The method optionally comprises a step E), wherein the by the captured pathogenic microorganisms extracellularly secreted and / or excreted stress responsive biomolecules are concentrated. By concentrating the extracellularly secreted or excreted stress responsive biomolecules 101 the concentration of the extracellularly secreted or excreted stress responsive biomolecules 101 increases. This may for example be performed by heating, in particular by evaporating at least a fraction of the supernatant of the captured pathogenic microorganisms and therein suspended extracellularly secreted or excreted stress responsive biomolecules. The method may further comprise a step D), wherein the by the captured pathogenic microorganisms extracellularly secreted stress responsive biomolecules are analysed, preferably by means of an imaging modality 106. Step D) may for example be performed by means of Raman spectroscopy, Surface-Enhanced Raman Spectroscopy (SERS) and / or biochemical analysis.

[0121] It will be clear that the invention is not limited to the exemplary embodiments which are illustrated and described here, but that countless variants are possible within the framework of the attached claims, which will be obvious to the person skilled in the art. In this case, it is conceivable for different inventive concepts and / or technical measures of the above-described variant embodiments to be completely or partly combined without departing from the inventive idea described in the attached claims.

[0122] The verb 'comprise' and its conjugations as used in this patent document are understood to mean not only 'comprise', but to also include the expressions 'contain', 'substantially contain', 'formed by' and conjugations thereof.

Claims

Claims1 . Method for processing at least one body fluid sample for pathogenic microorganism analysis, comprising the steps of:A) providing at least one body fluid sample,B) filtering the at least one body fluid sample through at least one catch filter which is configured for capturing at least a fraction of pathogenic microorganisms from the at least one body fluid sample if present in the body fluid sample, andC) applying at least one stress stimulus to at least part of the at least one catch filter, in particular to the fraction of pathogenic microorganisms, such that at least a fraction of the captured pathogenic microorganisms extracellularly secrete stress responsive biomolecules.

2. Method according to claim 1 , wherein the steps A) to C) are subsequent steps.

3. Method according to any of the preceding claims, wherein during step C) at least one chemical stress stimulus is applied to at least part of the at least one catch filter.

4. Method according to claim 3, wherein the chemical stress stimulus comprises at least one fluid and / or at least one chemical substance, preferably chosen from the group consisting of: water, demi-water, an acid, a base, an antimicrobial substance and / or a combination thereof.

5. Method according to any of the preceding claims, wherein during step C) at least one mechanical stress stimulus is applied to at least part of the at least one catch filter.

6. Method according to claim 5, wherein the mechanical stress stimulus is a mechanical force.

7. Method according to any of the preceding claims, wherein during step C) at least one electrical stress stimulus is applied to at least part of the at least one catch filter.

8. Method according to any of the preceding claims, wherein during step C) at least one stress stimulus is applied by a change in temperature and / or by electromagnetic radiation.

9. Method according to any of the preceding claims, comprising a step D) of analysing the by the captured pathogenic microorganisms extracellularly secreted stress responsive biomolecules.

10. Method according to claim 9, wherein step D) is performed by means of Raman Spectroscopy, Surface-Enhanced Raman Spectroscopy (SERS) and / or biochemical analysis.11 . Method according to any of the preceding claims, comprising a step of identifying at least one type of the captured pathogenic microorganism, in particular based on the extracellularly secreted stress responsive biomolecules.

12. Method according to any of the preceding claims, wherein at least part of the catch filter and / or at least part of the captured pathogenic microorganisms is provided in and / or dissolved in at least one fluid.

13. Method according to claim 12, comprising step E) of concentrating at least part of the at least one fluid such that the concentration of captured pathogenic microorganisms and / or the concentration of extracellularly secreted stress responsive biomolecules is increased.

14. Method according to claim 13, wherein step E) is performed by heating.

15. Method according to any of the preceding claims, comprising a step F) of filtering the at least one body fluid sample through at least one coarse filter which is configured for removing at least a fraction of particles from the body fluid sample, in particular particles which are larger than pathogenic microorganisms.

16. Method according to claim 15, wherein step F) is performed prior to step B).

17. Method according to any of the preceding claims, wherein at least a part of the catch filter is coated with nanoparticles, in particular SERS-active nanoparticles to form a SERS substrate.

18. Method according to any of the preceding claims, comprising a step G) of filtering at least one solution comprising nanoparticles, in particular SERS-active nanoparticles through the catch filter such that at least part of the catch filter will be coated with said nanoparticles.

19. Device for processing at least one body fluid sample for pathogenic microorganism analysis, comprising:- at least one housing, comprising: o at least one retaining structure for retaining at least one catch filter, and o at least one stress applicator for applying at least one stress stimulus to at least part of at least one catch filter retained in at least one retaining structure,- optionally, at least one catch filter configured to capture at least a fraction of pathogenic microorganisms from a body fluid sample, wherein at least one housing is a modular housing which is adjustable between:- a filtering configuration wherein at least one body fluid can be filtered through at least one catch filter retained in the at least one retaining structure; and- a processing configuration wherein at least one retaining structure is positioned such that at least part of at least one stress applicator is, directly or indirectly, in contact with at least part of at least one retaining structure and / or at least one catch filter retained in the at least one retaining structure.

20. Device according to claim 19, wherein at least one retaining structure is displaceable at least between the filtering configuration and the processing configuration.21 . Device according to any of the claims 19 to 20, wherein at least one stress applicator is a chemical stress applicator configured to apply at least one chemical stress stimulus to at least part of at least one catch filter retained in at least one retaining structure, in particular to the captured fraction of pathogenic microorganisms from a body fluid sample if present in the body fluid sample.

22. Device according to any of the claims 19 to 21 , wherein at least one stress applicator is an electrical stress applicator configured to apply at least one electrical stress stimulus to at least part of at least one catch filter retained in at least one retaining structure, in particular to the captured fraction of pathogenic microorganisms from a body fluid sample if present in the body fluid sample.

23. Device according to any of the claims 19 to 22, wherein at least one stress applicator is a mechanical stress applicator configured to apply at least one mechanical stress stimulus to at least a part of at least one catch filter retained in at least one retaining structure, in particular to the captured fraction of pathogenic microorganisms from a body fluid sample if present in the body fluid sample.

24. Device according to any of the claims 19 to 23, wherein at least one retaining structure is detachable connected to the housing.

25. Device according to any of the claims 19 to 24, wherein the housing further comprises at least one fluid reservoir for receiving at least one fluid, and wherein in the processing configuration at least one retaining structure is positioned adjacent to at least part of the at least one fluid reservoir such that fluid received in the fluid reservoir can get into contact with at least part of at least one catch filter retained in the at least one retaining structure.

26. Device according to claim 25, wherein in the processing configuration at least one catch filter retained in the retaining structure at least partially covers the fluid reservoir.

27. Device according to any of the claims 25 to 26, comprising at least one sealing member configured to couple the fluid reservoir and the retaining structurein a liquid-proof or liquid-tight manner, preferably at least in the processing configuration.

28. Device according to any of the claims 25 to 27, wherein the fluid reservoir is detachable connected to the housing.

29. Device according to any of the claims 25 to 28, wherein at least part of the at least one fluid reservoir comprises a translucent or transparent portion.

30. Device according to any of the claims 25 to 29, wherein the housing comprises at least one processing opening which is at least partially aligned with the fluid reservoir in the processing configuration for passaging fluid to the fluid reservoir in the processing configuration.31 . Device according to any of the claims 25 to 30, comprising at least one fluid applicator for applying fluid to the at least one fluid reservoir in the processing configuration.

32. Device according to claim 31 , wherein at least one fluid applicator comprises a solution comprising SERS-active nanoparticles to be filtered through the catch filter retained in the retaining structure, such that at least part of the catch filter will be coated with SERS-active nanoparticles thereby forming a SERS substrate.

33. Device according to claim 21 and any of the claims 31 to 32, wherein at least one chemical stress applicator is a fluid applicator comprising a fluid configured to apply a stress stimulus to the at least part of at least one catch filter retained in the at least one retaining structure, in particular to the captured fraction of pathogenic microorganisms from a body fluid sample if present in the body fluid sample.

34. Device according to any of the claims 31 to 33, wherein at least one fluid applicator is connectable to at least one processing opening for passaging fluid from the fluid applicator to the fluid reservoir in the processing configuration.

35. Device according to any of the claims 25 to 34, wherein the retaining structure comprises at least one inlet which is at least partially aligned with the fluid reservoir in the processing configuration for passaging fluid to the fluid reservoir in the processing configuration.

36. Device according to claim 34 and claim 35, wherein the processing opening and the inlet together form a fluid conduct in the processing configuration for guiding fluid to the fluid reservoir.

37. Device according to any of the claims 19 to 36, comprising at least one absorbing body for absorbing a fraction of fluid.

38. Device according to any of the claims 25 to 36 and claim 37, wherein at least absorbing body is provided adjacent to the fluid reservoir.

39. Device according to any of the claims 19 to 38, wherein the housing comprises at least one application opening which is at least partially aligned with the catch filter retained in the retaining structure in the filtering configuration for passaging at least one body fluid sample to the catch filter in the filtering configuration.

40. Device according to any of the claims 19 to 39, comprising at least one sample applicator for applying at least one body fluid sample to the catch filter at least in the filtering configuration.41 . Device according to claim 39 and claim 40, wherein the sample applicator is connected or connectable to at least one application opening.

42. Device according to any of the claims 19 to 41 , wherein the housing defines at least one accommodation space configured to at least partially accommodate at least one coarse filter which is configured for removing at least a fraction of particles from at least one body fluid sample, in particular particles which are larger than pathogenic microorganisms, and preferably wherein the at least oneaccommodation space is at least partially aligned with the retaining structure configured to retain at least one catch filter, at least in the filtering configuration.

43. Use of the sample processing device according to any of the claims 19 to 42 in a Raman Spectrometer for analysing extracellularly secreted stress responsive biomolecules of pathogenic microorganisms.

Images