Sample container, test system and test method for detecting pathogens and / or cells in a sample

Abstract

The invention relates to a sample container (10) for detecting pathogens and / or cells (11) in a sample, in particular a blood sample, with a sample processing unit (12), the sample container comprising: - at least one receiving region (13) of the sample processing unit (12) for receiving the sample, - at least one preparation region (14) of the sample processing unit (12) for preparing the sample, - at least one mixing region (15) of the sample processing unit (12) for thoroughly mixing the sample, the mixing region (15) comprising a mixing structure (16), and - at least one accumulating region (17) for accumulating the pathogens and / or cells (11) contained in the sample. The invention also relates to a test system (100) and a test method (200) for detecting pathogens and / or cells (11) in a sample.

Description

[0001] Sample container, test system and test procedure for the detection of pathogens and / or cells in a sample

[0002] Description

[0003] The invention relates to a sample container for detecting pathogens and / or cells in a sample, as well as a test system for detecting pathogens and / or cells in a sample and a corresponding test method.

[0004] Test systems for diagnosing diseases are already well-known in the state of the art. These include both laboratory-based diagnostic systems, particularly molecular diagnostic procedures and immunological tests, as well as point-of-care tests (PoC tests).

[0005] A PoC test is a near-patient diagnostic procedure that can be performed not in a central laboratory, but directly at the bedside or in a testing station by a physician or even a layperson. This has the advantage that results are immediately available to the patient. PoC test systems include, for example, pregnancy or coronavirus tests. In most cases, test strips are used for near-patient diagnostics. A sample (e.g., blood or urine) is applied to the test strip and transported along the strip by capillary action. The sample comes into contact with reagents that, in the presence of a target substance, trigger a chemical reaction or bond, resulting in a detectable color change (e.g., in the form of a stripe).

[0006] However, the disadvantage has emerged that PoC tests in the form of test strips have low accuracy and sensitivity. At low concentrations of a target substance, a negative result may be displayed even though the target substance (e.g., a pathogen) is present. This can be the case, for example, with tropical malaria infection, where parasite levels can be very low, particularly in pregnant women, asymptomatic carriers, and / or in the early stages of infection, so conventional test systems are often insufficient to reliably detect an infection.

[0007] Another disadvantage of known PoC tests is that they do not allow for real-time measurement of pathogen status. In a tropical malaria infection, for example, only a parasitic protein is detected, which circulates in the blood for many weeks. This prevents real-time measurement, which limits the possibility of monitoring drug effects, for example.

[0008] Furthermore, test strips are inevitably exposed to the environment, which means that the results can be influenced or distorted, particularly by high / low temperatures or humidity.

[0009] It is therefore the object of the present invention to at least partially remedy at least one of the disadvantages described above. In particular, the object of the present invention is to provide a sample container, a test system, and a test method for the detection of pathogens and / or cells in a sample, whereby pathogens and / or cells can be detected with particular precision in a cost-effective and time-saving manner with high sensitivity.

[0010] The above object is achieved by a sample container having the features of a first aspect of the invention, as well as by a test system having the features of a second aspect of the invention, and by a test method having the features of a third aspect of the invention. Further features and details of the invention emerge from the subclaims, the description, and the drawings. Features and details described in connection with the sample container according to the invention naturally also apply in connection with the test system according to the invention and / or in connection with the test method according to the invention, and vice versa, so that with regard to the disclosure of the individual aspects of the invention, reference is or can always be made reciprocally.According to a first aspect of the invention, a sample container for the detection of pathogens and / or cells in a sample, in particular a blood sample, is provided, comprising a sample processing unit comprising:.

[0011] - at least one receiving area of ​​the sample processing unit for receiving the sample,

[0012] - at least one processing area of ​​the sample processing unit for processing the sample,

[0013] - at least one mixing area of ​​the sample processing unit for mixing the sample, wherein the mixing area comprises a mixing structure, and

[0014] - at least one enrichment area for enriching the pathogens and / or cells contained in the sample.

[0015] A sample container can be understood, in particular, as a reaction vessel. The sample container can preferably be made at least partially from polypropylene and / or from cycloolefin copolymer (ethylene norbornene copolymer). The sample container can, in particular, have a conical shape and / or be produced by an injection molding process. Furthermore, the sample container can have walls, wherein an angle of the walls of the sample container preferably tapers towards the bottom. Furthermore, it can be provided that the sample container has a planar bottom. Furthermore, it is conceivable that the sample container has a circular bottom surface, in particular with a diameter between 0.1 μm and 10 mm, preferably between 0.5 μm and 1 mm, particularly preferably between 1 μm and 100 μm.

[0016] The pathogens can preferably comprise at least one malaria pathogen and / or at least one human immunodeficiency virus (HIV). Furthermore, the cells can comprise at least one erythrocyte and / or one lymphocyte, in particular a T helper cell.

[0017] The sample may, in particular, be a blood sample, preferably comprising capillary blood from a fingertip and / or venous whole blood. The sample may further comprise another body fluid such as urine, saliva, synovial fluid, cerebrospinal fluid, plasma, serum, tear fluid, sweat, lymph fluid, and / or intercellular fluid. The sample container may comprise a sample processing unit, which may comprise a receiving area, a processing area, and / or a mixing area.

[0018] A receiving area can be understood as an area of ​​the sample container into which the sample, in particular a blood sample, is placed. The addition of the sample into the sample container can preferably be done manually or automatically.

[0019] A preparation area can be understood as an area of ​​the sample container in which the pathogens and / or cells contained in the sample are prepared. The preparation area preferably comprises a reagent, in particular a lyophilized and / or dried reagent. The reagent can in particular comprise at least one lysis agent and / or at least one permeabilizing agent, and / or at least one enriching agent and / or at least one dye and / or at least one stabilizer. In particular, the pathogens and / or cells are prepared by bringing them into contact with the preferably dissolved reagent.

[0020] A lysis agent can be understood in particular as an agent for the almost complete destruction of outer and / or inner cell membranes of the pathogens and / or cells. A permeabilizing agent can be understood as an agent for creating temporary and / or permanent permeability of outer and / or inner cell membranes of the pathogens and / or cells. An enrichment agent can preferably comprise at least one antibody with a magnetic capture agent for capturing the pathogens and / or cells from the sample. A staining agent can preferably comprise at least one antibody with a labeling agent for labeling the pathogens and / or cells in the sample. Stabilizers can be understood as substances that maintain the stability and functionality of the pathogens and / or cells, and / or of an enrichment agent and / or a staining agent.

[0021] A mixing area can be understood to be an area of ​​the sample container in which the sample is thoroughly mixed. For this purpose, a mixing structure can be arranged in the mixing area of ​​the sample container. The mixing structure can in particular have at least one mixing element, preferably in the form of spheres. Furthermore, the mixing structure can be designed with at least one mixing element in the form of an elevation, in particular in the form of small wings and / or edges and / or hills, which are arranged on an inner wall of the sample container in the mixing area. The mixing of the sample can preferably take place by centrifugation and / or an eccentric movement. Mixing elements can make the liquid to be mixed move more vigorously. During mixing, the sample is initially pressed against an edge of the sample container by centrifugal force.The mixing elements prevent the sample from adhering completely to the edge of the sample container, thus promoting a more effective distribution of the sample and / or reagent components. They can direct a mixture, especially a liquid, into the area of ​​the mixing elements, which helps to mix the sample and / or reagent more evenly within the mixture. The mixing elements create more turbulence in the liquid flow. The mixing of the sample and / or reagent can thus be further enhanced and made more efficient by optimizing the flow behavior of the mixture, especially the liquid.

[0022] Furthermore, the sample container has an enrichment region. An enrichment region can be understood as a region of the sample container in which the pathogens and / or cells contained in the sample are concentrated. The enrichment region preferably has at least one enrichment agent and / or at least one dye. The enrichment agent can in particular have at least one antibody for enriching the pathogens and / or cells contained in the sample, wherein the at least one antibody comprises a magnetic capture agent, in particular a magnetic microparticle, for capturing the pathogens and / or cells in the sample. The dye can comprise at least one further antibody with a labeling agent, in particular a fluorescent labeling agent, for labeling the pathogens and / or cells in the sample.A sandwich structure can preferably be created using the enrichment agent, the dye, a pathogen and / or a cell.

[0023] Enrichment in the enrichment region can preferably be carried out via a magnet using magnetic force, in particular at a bottom of the sample container. The at least one enrichment agent and / or the at least one coloring agent can be arranged in the sample container, in particular in the form of a lyophilized and / or dried reagent, wherein the at least one enrichment agent and / or the at least one coloring agent are preferably pre-stored on an inner wall and / or a bottom of the sample container. In addition, the reagent can in particular comprise at least one lysis agent and / or a permeabilizing agent, and / or at least one stabilizer. Pre-storage can also save time, since no reagents need to be added, allowing pre-prepared batches of the sample containers to be used close to the patient outside of laboratories.

[0024] The receiving area, the processing area, the mixing area, and the enrichment area of ​​the sample container can preferably be present as a homogeneous area with smooth transitions. Furthermore, the receiving area, the processing area, the mixing area, and the enrichment area of ​​the sample container can be present as a single area, with the areas preferably completely overlapping.

[0025] Overall, this enables particularly precise and sensitive detection of cells and / or pathogens in a sample, especially a blood sample. The mixed structure enables particularly efficient mixing of the sample, optimizing the enrichment of the pathogens and / or cells contained in the sample and thus, in particular, increasing the sensitivity of the detection. This allows even low concentrations of pathogens and / or cells in the sample to be detected.

[0026] It can further be provided within the scope of the invention that the enrichment region comprises at least one enrichment agent and at least one dye, wherein the enrichment agent comprises at least one antibody with at least one magnetic capture agent, in particular a magnetic microparticle, for capturing the pathogens and / or cells in the sample, wherein the at least one dye comprises at least one other antibody with at least one labeling agent, in particular a fluorescent labeling agent, for labeling the pathogens and / or cells in the sample, wherein a sandwich structure can preferably be generated by the at least one enrichment agent and the at least one dye. In other words, the enrichment region can have at least one enrichment agent and / or at least one dye.The enrichment agent comprises, in particular, the at least one antibody with a magnetic capture agent for capturing the pathogens and / or cells from the sample. The staining agent preferably comprises the at least one other antibody with a labeling agent for labeling the pathogens and / or cells in the sample. A sandwich structure can preferably be created by the at least one enrichment agent and the at least one staining agent, as well as at least one pathogen and / or one cell.

[0027] The antibodies are preferably monoclonal antibodies that can specifically bind to a target structure on the surface of the pathogens and / or cells. The target structure can, in particular, be an epitope on an antigen on the surface of the pathogens and / or cells.

[0028] It can be provided that at least one antibody is bound to a magnetic capture agent and at least one other antibody is bound to a labeling agent.

[0029] The magnetic capture agent can in particular comprise a magnetic microparticle, preferably a Dynabead and / or a Turbobead. The magnetic microparticle can have a diameter of 0.05 pm to 20 pm, preferably 0.07 pm to 10 pm, particularly preferably 0.5 pm to 5 pm.

[0030] The labeling agent may comprise a fluorescent labeling agent, in particular a fluorescent dye. The fluorescent labeling agent can preferably be excited to emit a fluorescent signal by optical excitation, wherein the fluorescent signal is detectable by a fluorescence microscope. The optical excitation occurs in particular in the near-infrared range, preferably at wavelengths between 700 and 2000 nm, in particular between 700 and 900 nm.

[0031] The antibody bound to a magnetic capture agent and the antibody bound to a labeling agent can each bind to different epitopes on an antigen on a surface of the pathogens and / or cells. This can create a sandwich structure generated by two antibodies and a pathogen and / or a cell. This can achieve the advantage that, by applying a magnet to the enrichment area of ​​the sample container, enrichment of at least one antibody bound to at least one magnetic capture agent can occur by magnetic force, wherein the at least one antibody is additionally bound to a target structure on a surface of the pathogens and / or cells contained in the sample.Thus, the pathogens and / or cells contained in the sample can be enriched in the enrichment area of ​​the sample container and even low concentrations of pathogens and / or cells in the sample can be detected.

[0032] Because the pathogens and / or cells are additionally bound to at least one antibody containing a marker, the enriched pathogens and / or cells can be detected particularly precisely, for example, by optical excitation. The advantage of using fluorescent labeling is that even low concentrations of pathogens and / or cells can be detected accurately, cost-effectively, and in a time-saving manner. With optical excitation in a wavelength range between 700 and 2000 nm, the autofluorescence of biological material is particularly low, which allows for better signal quality when optically exciting the fluorescent marker to detect the pathogens and / or cells contained in the sample.

[0033] Within the scope of the present invention, it is conceivable that the sample comprises a sample volume between 1 pL and 1 mL, preferably between 10 and 100 pL, particularly preferably between 20 and 30 pL.

[0034] This allows pathogens and / or cells to be detected with high sensitivity even in larger sample volumes. Enrichment, in particular, allows detection of pathogens and / or cells at low concentrations even in larger sample volumes.

[0035] Furthermore, it is conceivable that the enrichment region is arranged close to the bottom within the sample container, wherein in particular at least one bottom of the sample container is transparent in optical quality.

[0036] "Near the bottom" can be understood in particular as meaning that the enrichment region extends preferably between 0 pm and 200 pm, preferably between 0 pm and 50 pm, and particularly preferably between 0 pm and 25 pm, above the bottom of the sample container. 0 pm means that the enrichment region begins directly at the bottom.

[0037] The base of the sample container can preferably be planar. Furthermore, the base of the sample container can be made of cycloolefin copolymer (ethylene norbornene copolymer), glass, polyethylene terephthalate, polymethyl methacrylate, and / or polycarbonate. The base of the sample container is preferably made of cycloolefin copolymer (ethylene norbornene copolymer). Cycloolefin copolymers have the advantage of exhibiting particularly good optical properties, such as high light transmittance and low autofluorescence.

[0038] The design of the base of the sample container as transparent in optical quality can be understood in particular to mean that the base has a particularly high light transmittance. This allows sufficient fluorescent light to penetrate the sample through the base of the sample container upon fluorescence irradiation to make the pathogens and / or cells contained in the sample visible through fluorescence excitation. Furthermore, the transparent design of the base of the sample container in optical quality can ensure that, upon fluorescence excitation, sufficient fluorescent light emanating from the sample can be detected by a sensor arranged at the base of the sample container and / or perceived by the user's eye.

[0039] Furthermore, the sample container can be designed to be completely transparent and of optical quality, preferably made of cycloolefin copolymer (ethylene norbornene copolymer). This allows sufficient fluorescent light emanating from the sample to be detected by a sensor located at the bottom of the sample container and / or perceived by the user's eye when fluorescent light is irradiated from the side into the sample container upon fluorescence excitation.

[0040] The sensors are preferably designed as photomultiplier tubes (PMTs) or charge-coupled devices (CCDs). The optical quality of the substrate allows for particularly uniform image brightness, high image contrast, and / or high resolution when creating an image by an evaluation unit. Furthermore, geometric distortion and optical imaging errors, such as spherical or chromatic aberration, coma, and astigmatism, can be minimized. The image created by the evaluation unit preferably reveals the pathogens and / or cells contained in the sample.

[0041] Overall, the advantage achieved is that the optically advantageous design of the base enables particularly precise and time-saving detection of pathogens and / or cells in a sample and avoids possible errors in visualization due to optical impairments.

[0042] Furthermore, it is conceivable that the processing area comprises at least one lysis agent and / or at least one permeabilization agent for lysing and / or permeabilizing the pathogens and / or cells.

[0043] The lysis agent can comprise, for example, Triton X100 and / or deoxycholate. The permeabilizing agent can preferably comprise saponin and / or ammonium chloride. Lysis can be understood, in particular, as the almost complete destruction of outer and / or inner cell membranes of the pathogens and / or cells. Permeabilization can be understood, in particular, as a process for creating a temporary and / or permanent permeability of outer and / or inner cell membranes of the pathogens and / or cells. Lysis and / or permeabilization each allow access to the internal structures of the pathogens and / or cells.

[0044] The advantage of lysis and / or permeabilization of the cells contained in the sample is that intracellular structures can also be detected. For example, pathogens such as parasites, bacteria or viruses can penetrate a so-called host cell, making exposure of the intracellular structures essential for the detection of these pathogens. Furthermore, lysis and / or permeabilization can expose the inner side of a cell membrane, allowing structures located there, such as specific proteins, to be detected. Lysis and / or permeabilization of the pathogens and / or cells contained in the sample can release biomolecular components of the pathogens and / or cells, such as proteins, nucleic acids or lipids, for particularly specific detection.

[0045] Overall, a particularly precise detection of pathogens and / or cells can be achieved, since specific components of the pathogens and / or cells can be detected.

[0046] The above object is further achieved by a second aspect of the invention. The second aspect of the invention comprises a test system for detecting pathogens and / or cells in a sample, in particular a blood sample, comprising:

[0047] - at least one sample container according to the first aspect of the invention, wherein the sample container comprises a sample comprising pathogens and / or cells,

[0048] - at least one mixing unit with a first sample holder for receiving the sample container, wherein the mixing unit is designed to mix at least the sample by means of movement,

[0049] - at least one evaluation unit with a second sample holder for receiving the sample container, wherein the evaluation unit is designed to enrich the pathogens and / or cells in the enrichment region of the sample container and to visualize and detect at least one pathogen enriched in the enrichment region and / or at least one cell enriched in the enrichment region in the sample.

[0050] The sample container can preferably additionally comprise a reagent, in particular one stored upstream. The reagent can preferably be in lyophilized and / or dried form. In particular, the reagent comprises at least one lysis agent and / or at least one permeabilizing agent, and / or an enrichment agent and / or at least one dye and / or at least one stabilizer. The at least one lysis agent can preferably comprise Triton X for the almost complete destruction of cell membranes of the pathogens and / or the cells. The at least one permeabilizing agent can in particular comprise saponin for creating temporary and / or permanent permeability of the pathogens and / or the cells. The at least one enrichment agent can preferably comprise at least one antibody with a magnetic capture agent for capturing and enriching the pathogens and / or cells contained in the sample.The at least one staining agent can preferably comprise at least one antibody with a labeling agent for labeling the pathogens and / or cells contained in the sample. The at least one stabilizer can preferably comprise polysaccharides for preserving the components of the sample and / or the reagent.

[0051] The mixing unit may, in particular, comprise an eccentric mixer. Additionally, the mixing unit may preferably comprise a rotary mixer and / or a vibration mixer.

[0052] An eccentric mixer can be understood as a mechanical device that mixes components of a sample and / or a reagent through an eccentric rotary motion.

[0053] Preferably, the mixing of the sample and / or reagent in the sample container can be achieved, in particular by means of an eccentric mixer, by an elliptical movement and / or a rotational movement, with the sample container also rotating. Alternatively or additionally, the mixing of the sample in the sample container can be achieved by alternating vibration of the center of the sample container.

[0054] Furthermore, mixing the sample and / or the reagent in the sample container, in particular by means of an eccentric mixer, can comprise activation of the sample and / or lysis and / or permeabilization of the sample. Activation of the sample can be understood in particular as the generation of a sandwich complex consisting of an enrichment agent and a dye as well as a pathogen and / or a cell. The enrichment agent preferably comprises at least one antibody with a magnetic capture agent, in particular a magnetic microparticle, for capturing the pathogens and / or cells in the sample. The dye in particular comprises at least one other antibody with a labeling agent, in particular a fluorescent labeling agent, for labeling the pathogens and / or cells in the sample.The at least two antibodies can each bind in particular to an epitope, preferably to different epitopes, on an antigen on a surface of the pathogens and / or cells, thus creating a sandwich structure. The mixing unit can comprise a first sample holder, preferably in the form of a clamping device and / or a clip device. Alternatively or additionally, the first sample holder can also be designed as a recess in the mixing unit, wherein the sample container can be inserted into the recess and held there during mixing, in particular by a pressure pin and / or pressing. The mixing of the sample in the mixing unit can in particular be automated.

[0055] Furthermore, the mixing unit can be designed to control the temperature of the sample within the sample container. Temperatures between 0 and 100 °C, preferably between 20 and 60 °C, and particularly preferably between 30 and 40 °C, are particularly preferred.

[0056] The evaluation unit can be understood, in particular, as a device designed to enrich and / or visualize and detect the pathogens and / or cells contained in the sample. For this purpose, the evaluation unit can preferably comprise a magnet for enriching the pathogens and / or cells contained in the sample using magnetic force, wherein, in particular, the pathogens and / or cells are each bound to at least one enrichment agent, wherein the enrichment agent comprises at least one antibody with a magnetic capture agent, in particular a magnetic microparticle.

[0057] Furthermore, the evaluation unit can have an image processing unit, in particular with a microscope, preferably with a fluorescence microscope, for visualizing and detecting the pathogens and / or cells contained in the sample. For this purpose, the pathogens and / or cells are preferably each bound to at least one dye, wherein the dye comprises at least one antibody with a marker, in particular a fluorescent marker. The visualization and detection of the pathogens and / or cells contained in the sample by the evaluation unit can preferably be carried out using fluorescence excitation of the pathogens and / or cells provided with a fluorescent marker. The evaluation unit can comprise a second sample holder, preferably in the form of a clamp device and / or a clip device.Alternatively or additionally, the second sample holder can also be designed as a recess in the evaluation unit, whereby the sample container can be inserted into the recess and held there during mixing, in particular by a pressure pin and / or pressing. The visualization and detection of the pathogens and / or cells contained in the sample can preferably be automated.

[0058] Overall, this results in the advantage that a particularly time-saving and cost-effective detection of pathogens and / or cells in a sample is possible, whereby even low concentrations of pathogens and / or cells can be precisely detected.

[0059] It can further be provided within the scope of the invention that the sample container in the processing area has at least one lysis agent and / or at least one permeabilization agent for lysing and / or permeabilizing the cells and / or pathogens, wherein the mixing of the sample in the at least one mixing unit comprises the lysis and / or permeabilization of the pathogens and / or cells.

[0060] The lysis agent may, for example, comprise Triton X100 and / or deoxycholate. The permeabilizing agent may preferably comprise saponin and / or ammonium chloride.

[0061] This offers the advantage of enabling particularly time-saving and precise detection of pathogens and / or cells in a sample. Lysis and / or permeabilization of the pathogens and / or cells allows specific biomolecular structures of the pathogens and / or cells to be released, thus ensuring precise detection.

[0062] Furthermore, it can be provided within the scope of the invention that the evaluation unit comprises at least one magnet, wherein the evaluation unit is designed to enrich at least one of the pathogens and / or at least one of the cells by means of the magnet in the enrichment region of the sample container, wherein the at least one pathogen and / or the at least one cell is bound to at least one antibody with a magnetic capture agent, in particular a magnetic microparticle, wherein the enrichment region is arranged in particular close to the bottom within the sample container.

[0063] The at least one magnet can in particular be a ring magnet and / or a cylindrical magnet. The at least one magnet can preferably be designed as a neodymium magnet and / or as a switchable electromagnet and / or as a permanent magnet array. Furthermore, it can be provided that the at least one magnet is arranged in particular below the second sample holder in the evaluation unit. This preferably allows enrichment to take place in the enrichment region near the bottom of the sample container. Near the bottom can be understood to mean that the enrichment region preferably extends between 0 pm and 200 pm, preferably between 0 pm and 50 pm, particularly preferably between 0 pm and 25 pm, above the bottom of the sample container. 0 pm means that the enrichment region begins directly at the bottom.

[0064] Furthermore, it can be provided that the enrichment of the pathogens and / or cells contained in the sample takes place using antibodies with a bound magnetic capture agent, in particular with a bound magnetic microparticle. The antibody can bind specifically to a target structure on a surface of the pathogens and / or cells. By inserting the sample container into the second sample holder of the evaluation unit, the pathogens and / or cells provided with a magnetic capture agent can be enriched by the magnet via magnetic force in the enrichment area of ​​the sample container.

[0065] This can provide the advantage that the pathogens and / or cells can be detected in a sample with particularly high sensitivity by enrichment in the enrichment area of ​​the sample container.

[0066] Within the scope of the present invention, it is further conceivable that the evaluation unit is designed to visualize and detect the pathogens and / or cells through a base of the sample container in the enrichment region of the sample container, wherein the base is preferably transparent in optical quality, and / or wherein the evaluation unit is designed to detect individual pathogens and / or individual cells in the sample in the enrichment region of the sample container. The design of the base of the sample container as transparent in optical quality can be understood in particular to mean that the base has a particularly high light transmittance. As a result, when fluorescent light is irradiated through the base of the sample container, sufficient fluorescent light can reach the sample to make the pathogens and / or cells contained in the sample visible through fluorescence excitation.Furthermore, the transparent design of the bottom of the sample container in optical quality can ensure that, upon fluorescence excitation, sufficient fluorescent light emanating from the sample can be detected by a sensor arranged at the bottom of the sample container and / or perceived by the eye of a user.

[0067] Furthermore, the sample container can be designed to be almost completely transparent and of optical quality, preferably made of cycloolefin copolymer (ethylene norbornene copolymer). This allows sufficient fluorescent light emanating from the sample to be detected by a sensor located at the bottom of the sample container and / or perceived by the user's eye when fluorescent light is irradiated into the sample container from the side upon fluorescence excitation.

[0068] The sensors can preferably be designed as photomultiplier tubes (PMT) or charge-coupled devices (CCD).

[0069] The optical quality of the base allows for particularly uniform image brightness, high image contrast, and / or high resolution. Furthermore, geometric distortion and optical aberrations such as spherical or chromatic aberration, coma, and astigmatism can be minimized.

[0070] The visualization and detection of the individual pathogens and / or individual cells in the sample in the enrichment region of the sample container can be achieved by at least one staining agent. The at least one staining agent preferably comprises at least one antibody with a bound labeling agent, in particular a fluorescent labeling agent. The at least one antibody binds specifically to an epitope on an antigen on a surface of the pathogens and / or cells contained in the sample. The enrichment of the pathogens and / or cells contained in the sample in an enrichment region of the sample container can be achieved by at least one enrichment agent comprising at least one further antibody with a bound magnetic capture agent, in particular a magnetic microparticle. The at least one antibody can preferably bind specifically to one, in particular a different, epitope on an antigen on a surface of the pathogens and / or cells.By attaching a magnet, particularly in the evaluation unit, to the enrichment area, the pathogens and / or cells are enriched in the enrichment area.

[0071] By using a staining agent comprising a labeling agent, preferably a fluorescent labeling agent, individual enriched pathogens and / or cells can be visualized and detected, in particular by optical excitation and by means of a microscope, preferably a fluorescence microscope.

[0072] Because the pathogens and / or cells can be enriched in an enrichment area of ​​the sample container and then visualized and detected, the pathogens and / or cells in the sample can be detected with particularly high sensitivity. The ability to detect individual pathogens and / or cells means that even low concentrations of pathogens and / or cells in a sample can be detected. This can prevent the transmission of diseases, as even asymptomatic carriers and / or pregnant women with a very low pathogen and / or cell load can be detected and treated. Furthermore, the detection of individual pathogens and / or cells can be used to monitor medication, as a decrease or increase in the number of pathogens and / or cells can be precisely detected.

[0073] Furthermore, it is conceivable that the evaluation unit comprises an image processing unit, wherein the image processing unit is designed to visualize the pathogens and / or cells individually, wherein the image processing unit in particular comprises a, preferably inverse and / or automated, fluorescence microscope which is designed to visualize the pathogens and / or cells as fluorescent units.

[0074] The image processing unit may preferably comprise hardware and / or software, wherein the image processing unit is particularly designed to analyze, process and interpret image data of the enrichment region of the sample container.

[0075] The image processing unit may preferably comprise a fluorescence microscope, in particular an inverse and / or automated one, wherein the fluorescence microscope may have an optical magnification unit.

[0076] When using a fluorescence microscope, the pathogens and / or cells bound by a staining agent comprising at least one antibody with at least one bound fluorescent labeling agent can be detected by fluorescence excitation after enrichment in the enrichment region of the sample container.

[0077] The image processing unit, preferably the fluorescence microscope, can in particular comprise an objective lens and / or a light source. The objective lens can preferably be arranged below the sample container, wherein the light source can be arranged above or to the side of the sample container and / or below the sample container. The light source can generate radiation that is preferably in the near-infrared range and in particular comprises wavelengths from 700 to 2000 nm, preferably between 700 and 900 nm.

[0078] The image processing unit, preferably the fluorescence microscope, can in particular be designed to generate a series of images (Z-stack series) created by focusing on different planes of the sample in the sample container. Each image in a Z-stack series can show a specific section through the sample in the Z-axis (the vertical axis). The image processing unit can be designed to create 10 to 50 images, preferably 15 to 40 images, particularly preferably 20 to 30 images in different planes of the sample. The image processing unit can further be designed to create a single image from a Z-stack series by means of a maximum projection. The pathogens and / or cells contained in the sample are preferably visible in the image.

[0079] This offers the advantage of enabling particularly precise detection of pathogens and / or cells in a sample in a cost-effective and time-saving manner. Fluorescence detection also offers the advantage over conventional detection methods, such as Giemsa staining, of achieving higher sensitivity and specificity. Furthermore, waiting times can be reduced, as only very short incubation times need to be considered.

[0080] It can further be provided within the scope of the invention that the evaluation unit comprises a computing unit which is designed to quantify the at least one visualized pathogen and / or the at least one visualized cell and / or to carry out pattern recognition.

[0081] The computing unit can be designed to be mobile or permanently connected to the evaluation unit.

[0082] The computing unit can preferably comprise self-learning software. The computing unit can further be configured to determine a number of visualized pathogens and / or cells. Additionally or alternatively, the computing unit can be configured to perform pattern recognition, in particular by performing the following steps:

[0083] Performing a maximum projection of a Z-stack series to generate a single image of the enrichment area of ​​the sample container, wherein the Z-stack series is created by an image processing unit of the evaluation unit,

[0084] - Adjusting, in particular increasing, the contrast of the image,

[0085] Reducing the background of the image,

[0086] Detecting a pattern of visualized pathogens and / or cells in the image, wherein the image comprises in particular 1 to 20 megapixels, preferably 1 to 10 megapixels, particularly preferably 1 to 5 megapixels.

[0087] During pattern recognition, size minimization can also be performed, whereby in particular all objects visible in the image that are outside the desired size, preferably larger or smaller than 4 pixels, and / or outside the desired shape, preferably a shape of 2 x 2 pixels, are sorted out. The objects visible in the image can preferably include the pathogens and / or cells contained in the sample. This has the advantage of ensuring particularly precise detection of the pathogens and / or cells in a sample with high sensitivity. By precisely determining a number of pathogens and / or cells in the sample, disease progression can be determined, asymptomatic carriers with a low pathogen and / or cell load can be identified and treated, and drug resistance can be detected.Performing pattern recognition offers the advantage of a particularly time-saving and cost-effective detection of pathogens and / or cells in a sample.

[0088] Within the scope of the present invention, it is also conceivable that the evaluation unit

[0089] Communication interface designed to process analysis data of the

[0090] Evaluation unit to a mobile device and / or a database, in particular a

[0091] Data cloud, and / or wherein the evaluation unit comprises a display device, in particular a display, which is designed to display analysis data of the

[0092] Evaluation unit to display to a user.

[0093] The analysis data can preferably include a number of pathogens and / or cells in the sample and / or pattern recognition data. Furthermore, the analysis data can include personal data, preferably master data and / or medical data. The master data can, in particular, include a person's name, date of birth, and / or gender. Medical data can, in particular, include a person's medical history and / or medical reports.

[0094] It can further be provided that the analysis data contain information on the detection of pathogens and / or cells for a specific person, whereby, preferably after a new detection, a comparison of the number of pathogens and / or cells in a sample can be performed. This allows, for example, drug resistance to be detected early.

[0095] The communication interface can in particular be designed to transmit the analysis data in a wired and / or wireless manner, preferably via Bluetooth and / or WLAN (Wireless Local Area Network).

[0096] The mobile device can preferably be a tablet, a smartphone, or a smartwatch. The database can preferably comprise a data cloud, whereby analysis data can be retrieved via the data cloud, which contains, in particular, anonymized information about detected pathogens and / or cells in a sample (positive test result), in particular in connection with a location of detection. This allows prevalence maps to be created, whereby locations with a higher detection rate can be supplied to medical personnel and medications.

[0097] This offers the advantage of using a database to localize and contain the spread of disease. Furthermore, the patient or physician can read the results directly on a mobile device or display, saving time by eliminating the need to analyze complex laboratory reports.

[0098] The above object is further achieved by a third aspect of the invention. The third aspect of the invention comprises a test method for detecting pathogens and / or cells in a sample, in particular a blood sample, preferably using a test system according to the second aspect of the invention, comprising the following steps:

[0099] Introducing a sample, in particular a blood sample, into a receiving area of ​​a sample preparation unit of a sample container, wherein the sample comprises pathogens and / or cells,

[0100] - Preparation of the sample in a preparation area of ​​the sample preparation unit of the sample container,

[0101] Mixing the sample in a mixing area of ​​the sample preparation unit of the sample container by placing the sample container into a mixing unit,

[0102] - Enrichment of the pathogens and / or cells in the sample in an enrichment area of ​​the sample container by placing the sample container in an evaluation unit,

[0103] - Evaluation of the enrichment area of ​​the sample container by the evaluation unit, whereby the pathogens and / or cells in the sample are visualized and detected by the evaluation unit.

[0104] The described test method can preferably be performed by medical personnel, laypersons (medically untrained personnel), or by a patient. Furthermore, the test method can be performed automatically or manually. Furthermore, the evaluation of the enrichment area of ​​the sample container can include real-time quantification of pathogens and / or cells in a sample. Furthermore, the evaluation of the enrichment area of ​​the sample container can include visualization and detection of individual pathogens and / or cells.

[0105] Furthermore, a reagent, in particular a lyophilized and / or dried one, can be stored in the sample container, wherein the reagent can in particular comprise at least one lysis agent and / or at least one permeabilizing agent, and / or at least one enriching agent and / or at least one dye and / or at least one stabilizer. The lysis agent can in particular comprise Triton X100 and / or deoxycholate. The permeabilizing agent can preferably comprise saponin and / or ammonium chloride. The enriching agent can preferably comprise at least one antibody that can bind pathogens and / or cells in the sample, wherein the at least one antibody can comprise a magnetic capture agent, preferably a magnetic microparticle.The staining agent can preferably comprise another antibody with a labeling agent, preferably a fluorescent labeling agent, wherein the labeling agent can, in particular, label the pathogens and / or cells in the sample. The two antibodies and a pathogen and / or a cell can preferably form a sandwich structure. The stabilizer can, in particular, comprise polysaccharides, preferably dextran.

[0106] It may be provided that a sample to be examined, in particular a blood sample, is added to the reagent contained in the sample container, in particular a lyophilized and / or dried reagent. Furthermore, it is conceivable that the sample comprising the pathogens and / or cells is brought into contact with the reagent.

[0107] Subsequently, the sample container containing the reagent, in particular the lyophilized and / or dried reagent, and the sample can preferably be placed into the first sample holder of the mixing unit, in particular comprising an eccentric mixer. The mixing unit can preferably mix the sample thoroughly, whereby the mixing can, in particular, result in the lysis and / or permeabilization of the pathogens and / or cells in the sample and / or an enrichment of the pathogens and / or cells in the sample in the enrichment area of ​​the sample container and / or a labeling of the pathogens and / or cells in the sample.

[0108] It is further conceivable that the sample container is subsequently placed in a second sample holder of the evaluation unit. The evaluation unit can preferably comprise a magnet and / or a microscope, in particular a fluorescence microscope. The pathogens and / or cells are preferably bound to at least one enrichment agent with at least one antibody, which can be bound to a magnetic capture agent. As a result, the pathogens and / or cells in the sample can be enriched in the enrichment region by means of the magnet through magnetic force. The pathogens and / or cells are furthermore bound in particular to at least one staining agent with at least one other antibody, which can be bound to a labeling agent, and can be visualized and detected by means of the microscope.

[0109] This offers the advantage of enabling precise detection of pathogens and / or cells in a sample in a simple, cost-effective manner and with a time-saving approach. In particular, the sensitivity of the test procedure can be improved by enriching the pathogens and / or cells.

[0110] Within the scope of the invention, it can further be provided that the preparation of the sample comprises a lysis and / or a permeabilization of the pathogens and / or cells, and / or wherein the enrichment of the pathogens and / or cells in the enrichment region of the sample comprises a magnetic capture and / or a fluorescent label, and / or wherein the evaluation of the enrichment region comprises a quantification of the pathogens and / or cells or a pattern recognition, wherein the enrichment region is arranged in particular close to the bottom within the sample container and the evaluation of the enrichment region takes place in particular through a bottom of the sample container, wherein the bottom is transparent in optical quality.

[0111] The lysis agent can comprise, for example, Triton X100 and / or deoxycholate. The permeabilizing agent can preferably comprise saponin and / or ammonium chloride. The lysis and / or permeabilization each allow access to the internal structures of the pathogens and / or cells. For the enrichment of the pathogens and / or cells contained in the sample in the enrichment region, the pathogens and / or cells can be bound to at least one enrichment agent. The at least one enrichment agent preferably comprises at least one antibody with a magnetic capture agent, preferably a magnetic microparticle. By applying a magnet, the pathogens and / or cells provided with the magnetic capture agent can be concentrated in the enrichment region of the sample container.

[0112] The pathogens and / or cells contained in the sample can preferably be additionally bound to at least one staining agent. The at least one staining agent preferably comprises a further antibody with a labeling agent, preferably a fluorescent labeling agent.

[0113] The at least two antibodies can each bind to a different epitope on an antigen on a surface of the pathogens and / or cells. Thus, a sandwich structure can be created by the at least one enrichment agent and the at least one staining agent, as well as a pathogen and / or a cell. The sandwich structure can preferably be enriched in the enrichment area of ​​the sample container, in particular by a magnet.

[0114] The enriched pathogens and / or cells can preferably be visualized and detected individually, in particular by a fluorescence microscope.

[0115] Overall, the advantage can be achieved that the sensitivity and specificity of the detection of pathogens and / or cells in a sample is further improved, since biomolecular structures of pathogens and / or cells can also be examined and even low concentrations of pathogens and / or cells can be detected via enrichment.

[0116] Furthermore, within the scope of the invention, it is conceivable that the pathogens comprise at least one malaria pathogen, in particular Plasmodium falciparum and / or Plasmodium vivax and / or Plasmodium ovale and / or Plasmodium knowlesi and / or Plasmodium malariae, and / or the cells comprise at least one erythrocyte. This results in the advantage that the test method can be used to detect malaria. Malaria is a deadly disease that is particularly prevalent in areas where no laboratory diagnostics are available. Thus, cost-effective and time-saving detection of malaria pathogens, which can also be performed by laypersons (medically untrained personnel) and / or by the patient themselves, is essential for controlling malaria.

[0117] Furthermore, it is conceivable that the pathogens comprise at least one HIV virus and / or the cells comprise at least one lymphocyte, preferably a T helper cell, particularly preferably a CD4 T cell and / or CD8 T cell.

[0118] It can be provided that when the evaluation unit evaluates the enrichment region of the sample container, a decrease in the number of T helper cells, in particular CD4 T cells and / or CD8 T cells, can be detected. It is conceivable that the enrichment of the T helper cells takes place by means of at least one enrichment agent comprising at least antibodies with at least one bound magnetic capture agent, in particular at least one magnetic microparticle. In this case, the at least one antibody can bind with the magnetic capture agent to an epitope on an antigen on the surface of the T helper cells. Detection of the enriched T helper cells can take place via a staining agent, wherein the staining agent preferably comprises another antibody with at least one bound marker, in particular at least one fluorescent marker.The at least one other antibody can bind with the marker to one, in particular a different, epitope on an antigen on the surface of the T helper cells. This allows a sandwich complex comprising the two antibodies and the T helper cell to be generated, wherein the sandwich complex can be enriched in the enrichment area of ​​the sample container with a magnet and visualized and detected using an image processing unit, in particular comprising a fluorescence microscope.

[0119] The advantage of this test procedure is that it can detect an outbreak of AIDS, which can be triggered by HIV viruses that infect T helper cells, by detecting T helper cells in a sample with particularly high sensitivity and, in particular, by monitoring the number of T helper cells in the sample.

[0120] Furthermore, it can be provided within the scope of the invention that in particular the disease is a disease caused by pathogens, in particular fungi, viruses and / or bacteria, and / or pathogens, in particular at least one from the group of Adenoviruses, Amylostoma, Ascaris, Babesia, Bacillus anthracis, Bordetella pertussis, Bordetella parapertussis, Borrelia recurrentis, Brucella sp., Campylobacter sp., Cestoda, Chlamydia psittaci, Clostridium botulinum, Corynebacterium diphtheriae, Coxiella burnetii, human pathogenic Cryptosporidium sp., Ebola virus, Echinococcus multilocularis, Echinococcus granulosus, Escherichia coli, enterohemorrhagic strains (EHEC), Eucestoda, Francisella tularensis, TBE- irus, yellow fever virus, Giardia lamblia, Haemophilus influenzae, Hantaviruses, Hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus, hepatitis E virus, influenza viruses, Lassa virus, Legionella sp., human pathogenic Leptospira sp., Listeria monocytogenes, Marburgvirus, Measles virus, Mumps virus, Mycobacterium leprae, Mycobacterium tuberculosis / africanum, Mycobacterium bovis, Neisseria meningitidis, Norwalk-like virus, Poliovirus, Pseudomonas aeruginosa, Rabies virus, Rickettsia prowazekii, Rotavirus, Rubella virus, Salmonella Paratyphi, Salmonella Typhi, Schistosoma, Shigella sp., Taenia saginata, Taenia solium, Trichiuris, Trypanosoma, Trypanosoma brucei, Trypanosoma congolense, Trypanosoma vivax, Trichinella spiralis, Varicella-Zoster virus, Vibrio cholerae 1 and 139, Yersinia enterocolitica, Yersinia pestis, Treponema pallidum, HIV, Echinococcus sp., Plasmodium sp., Toxoplasma gondii, Streptococcus pneumoniae and / or Streptococcus aureus.

[0121] Brucella sp. (species) may include Brucella abortus, Brucella melitensis, Brucella suis, and / or Brucella canis. Campylobacter sp. may include Campylobacter jejuni and / or Campylobacter coli. Legionella sp. may include Legionella pneumophila, Legionella micdadei, and / or Legionella longbeachae. Shigella sp. may include Shigella dysenteriae, Shigella flexneri, and / or Shigella boydii. Echinococcus sp. may include Echinococcus granulosus, Echinococcus multiocularis, Echinococcus vogeli, and / or Echinococcus oligarthrus. Plasmodium sp. may include Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale, Plasmodium malariae, and / or Plasmodium knowlesi. This has the advantage that the test procedure can detect a wide range of pathogens precisely and specifically, making the procedure versatile and suitable for diagnosing many diseases.

[0122] Further advantages, features, and details of the invention will become apparent from the following description, in which several embodiments of the invention are described in detail with reference to the drawings. The features mentioned in the claims and in the description may be essential to the invention individually or in any combination. The invention is explained in more detail below with reference to the accompanying drawings. These show schematically:

[0123] Figure 1 Representation of a sample container for the detection of pathogens and / or cells in a sample,

[0124] Figure 2 Representation of a sandwich structure comprising pathogens and / or cells to be detected in a sample,

[0125] Figure 3 schematic representation of the test system for the detection of pathogens and / or cells in a sample,

[0126] Figure 4 Representation of the enriched pathogens and / or cells in an enrichment area of ​​the sample container,

[0127] Figure 5 schematic representation of the test procedure for the detection of pathogens and / or cells in a sample.

[0128] In the following figures, identical reference numerals are used for the same technical features, even for different embodiments.

[0129] Fig. 1 shows a sample container 10 with a sample processing unit 12, wherein the sample processing unit 12 comprises a receiving area 13, a preparation area 14, and / or a mixing area 15. The receiving area 13 can be understood as an area into which a sample is introduced into the sample container 10. The preparation area 14 can be understood as the area in which the sample is prepared. The mixing area 15 can be understood as an area in which the sample is mixed, preferably by movement. For this purpose, the mixing area 15 can preferably have a mixing structure 16. The mixing structure 16 can preferably comprise mixing elements 16a, in particular spheres.Alternatively or additionally, the mixing structure 16 can be designed in the form of mixing elements 16b, in particular in the form of elevations as wings and / or edges and / or hills, which are arranged on an inner wall of the sample container 10 in the mixing region 15.

[0130] Furthermore, the sample container 10 comprises an enrichment region 17. The enrichment region 17 can be understood as a region in which the pathogens and / or cells 11 contained in the sample are concentrated.

[0131] The receiving area 13, the processing area 14, the mixing area 15 and the enrichment area 17 can also form a single area, wherein the areas 13, 14, 15 and 17 preferably completely overlap.

[0132] The sample container 10 can be understood as a reaction vessel. The sample container 10 is preferably made at least partially of polypropylene and / or cycloolefin copolymer (ethylene norbornene copolymer). Furthermore, the sample container 10 preferably has a conical shape. The sample container 10 comprises, in particular, walls, wherein an angle of the walls preferably tapers downwards.

[0133] A base 24 of the sample container 10 is preferably planar. Furthermore, the base 24 of the sample container 10 is particularly transparent and optically transparent. This enables easy detection of the pathogens and / or cells 11 contained in the sample through the base 24 of the sample container 10.

[0134] Fig. 2 shows a sandwich structure 18, which is formed in particular by an enrichment agent 19 and a staining agent 20 as well as a pathogen and / or a cell 11. The enrichment agent 19 preferably comprises an antibody 21a. The staining agent 20 preferably has another antibody 21b. One antibody 21a preferably comprises a magnetic capture agent 22, wherein the magnetic capture agent 22 can be designed in particular as a magnetic microparticle. The other antibody 21b preferably comprises a labeling agent 23, wherein the labeling agent 23 can preferably be designed as a fluorescent labeling agent. One antibody 21a can bind to a target structure on the surface of the pathogen and / or the cell 11, while the other antibody 21b can bind to a, preferably different, target structure on the surface of the pathogen and / or the cell 11.Thus, a sandwich structure 18 can be created by the enrichment agent 19, the dye 20 and a pathogen and / or a cell 11.

[0135] Fig. 3 shows a test system 100 for the detection of pathogens and / or cells 11 in a sample. The test system 100 comprises a sample container 10 containing a sample with the pathogens and / or cells 11. The test system 100 further comprises a mixing unit 101 with a first sample holder 102, wherein the first sample holder 102 is designed to receive the sample container 10. The mixing unit 101 is designed to mix a reagent stored upstream in the sample container 10 and the sample in the sample container 10. The reagent preferably comprises at least one lysis agent and / or at least one permeabilizing agent, and / or at least one enriching agent 19 and / or at least one coloring agent 20 and / or at least one stabilizer. The mixing unit 101 can preferably comprise an eccentric mixer. In addition, the mixing unit 101 may in particular comprise a rotary mixer and / or a vibration mixer.

[0136] Furthermore, the test system 100 has an evaluation unit 103, which is designed to enrich the pathogens and / or cells 11 in an enrichment region 17 of the sample container 10 and to visualize and detect at least one pathogen 11 enriched in the enrichment region 17 and / or at least one cell 11 enriched in the enrichment region in the sample. The evaluation unit 103 preferably comprises a magnet 105 for enriching the pathogens and / or cells 11 contained in the sample, wherein the pathogens and / or cells 11 are present in particular in a sandwich structure 18, an enrichment agent 19, and a dye 20. The enrichment agent 19 preferably comprises at least one antibody 21a with at least one magnetic capture agent 22, in particular a magnetic microparticle.The staining agent 20 preferably comprises another antibody 21b with a marker 23, in particular a fluorescent marker. The magnet 105 can preferably be arranged below the second sample holder 104, thus ensuring enrichment of the pathogens and / or cells 11 contained in the sample in the enrichment region 17 of the sample container 10. The enrichment region 17 is arranged near the bottom 24, in particular directly at the bottom 24, of the sample container 10.

[0137] Furthermore, the evaluation unit 103 can have an image processing unit 106, preferably comprising a fluorescence microscope 107. Using the image processing unit 106, the pathogens and / or cells 11 in the sample can be individually visualized and detected, preferably as fluorescent units.

[0138] Furthermore, the evaluation unit 103 can comprise a computing unit 108. The computing unit 108 can be configured to quantify the at least one pathogen 11 visualized by the image processing unit 106 and / or the at least one cell 11 visualized by the image processing unit 106. Alternatively or additionally, the computing unit can be configured to perform pattern recognition, wherein a Z-stack series recorded by the image processing unit 106 is processed into a single image using a maximum projection, and pattern recognition of the visualized and detected pathogens and / or cells is performed based on this image.In particular, size minimization can be carried out during pattern recognition, whereby preferably all objects visible in the image outside the desired size, preferably larger or smaller than 4 pixels, and / or outside the desired shape, preferably a shape of 2 x 2 pixels, are sorted out.

[0139] Furthermore, the evaluation unit 103 can have a communication interface 109, which is preferably designed to transmit analysis data to a mobile device 110, in particular a smartphone, and / or to a database 111, in particular a data cloud, in a wired and / or wireless manner, preferably via Bluetooth and / or WLAN.

[0140] The evaluation unit 103 may further comprise a display device 112, in particular a display, for displaying analysis data to a user.

[0141] Fig. 4 shows a top view of a surface of the base 24 of the sample container 10, wherein individual pathogens and / or individual cells 11 have been visualized in an enrichment region 17 of the sample container 10, in particular as fluorescent units, and individual detection of the pathogens and / or cells 11 is possible. Based on the individual detection, in particular, a quantification of the pathogens and / or cells 11 and / or pattern recognition can be performed. The arrangement of the pathogens and / or cells 11 preferably resembles a starry sky.

[0142] Fig. 5 shows a test method 200 for detecting pathogens and / or cells 11 in a sample. The test method 200 comprises, in a first step, introducing 201 a sample, in particular a blood sample, into a receiving area 13 of a sample preparation unit 12 of a sample container 10, wherein the sample comprises pathogens and / or cells 11. A further step of the test method 200 comprises preparing 202 the sample in a processing area 14 of the sample preparation unit 12 of the sample container 10. In a further step, the test method 200 comprises mixing 203 the sample in a mixing area 15 of the sample preparation unit 12 of the sample container 10 by introducing the sample container 10 into a mixing unit 101.A further step of the test method 200 includes the enrichment 204 of the pathogens and / or cells 11 in the sample in an enrichment region 17 of the sample container 10 by introducing the sample container 10 into an evaluation unit 103. A final step of the test method 200 comprises an evaluation 205 of the enrichment region 17 of the sample container 10 by the evaluation unit 103, wherein the pathogens and / or cells 11 in the sample are visualized and detected by means of the evaluation unit 103.

[0143] It can preferably be provided that a reagent, in particular a lyophilized and / or dried one, is stored upstream in the sample container 10. The reagent preferably comprises at least one lysis agent and / or at least one permeabilizing agent, and / or at least one enrichment agent 19 and / or at least one coloring agent 20, and / or at least one stabilizer. The sample, in particular a blood sample, can then be added to the reagent. The enrichment agent 19 can preferably comprise an antibody 21a bound to a magnetic capture agent 22, in particular a magnetic microparticle 19. The coloring agent 20 can preferably comprise an antibody 21b with a labeling agent 23, in particular a fluorescent labeling agent. The stabilizer can in particular comprise polysaccharides, preferably dextran, to maintain the functionality of the sample and / or the reagent.The sample container 10 with the, in particular lyophilized and / or dried, reagent and the sample can preferably be placed in a first sample holder 102 of the mixing unit 101, in particular comprising an eccentric mixer. The mixing unit 101 can preferably mix the sample and / or the reagent, wherein the mixing can in particular result in the lysis and / or permeabilization of the pathogens and / or cells 11 in a sample and / or an enrichment of the pathogens and / or cells 11 in the sample in the enrichment region 17 of the sample container 10 and / or a marking of the pathogens and / or cells 11 in the sample.

[0144] It is further conceivable that the sample container 10 is subsequently placed in a second sample holder 104 of the evaluation unit 103. The evaluation unit 103 can preferably comprise a magnet 105 and / or a fluorescence microscope 107. The pathogens and / or cells 11 are preferably bound to an enrichment agent 19 containing an antibody 21a, wherein the antibody 21a is in particular bound to a magnetic capture agent 22. The magnet 105 can thus enrich the pathogens and / or cells 11 in the enrichment region 17 by means of magnetic force. The enriched pathogens and / or cells 11 are in particular additionally bound to a staining agent 20 containing an antibody 21b, wherein the antibody 21b is in particular bound to a labeling agent 23, in particular a fluorescent labeling agent. Thus, the pathogens and / or cells 11 can be visualized and detected using the fluorescence microscope 107.

[0145] This enables particularly precise detection of cells and / or pathogens 11 in a sample, in particular a blood sample, with high sensitivity, whereby even individual pathogens and / or individual cells 11 can be detected. Thus, even very low concentrations of pathogens and / or individual cells 11 in a sample can be detected, and an increase and / or decrease in an exact number of pathogens and / or individual cells 11 in a sample can be determined.

[0146] The above description of the figures describes the present invention by way of examples. Of course, individual features of the embodiments can be freely combined with one another, provided they are technically feasible, without departing from the scope of the invention. Reference symbols

[0147] Sample container

[0148] Pathogens and / or cells Sample processing unit Collection area Preparation area Mixing area Mixing structure

[0149] Enrichment area Sandwich structure Enrichment agent Staining agent a Antibody b Antibody

[0150] Magnetic catching device

[0151] Marking agents

[0152] Soil 0 Test system 1 Mixing unit 2 First sample holder 3 Evaluation unit 4 Second sample holder 5 Magnet 6 Image processing unit 7 Microscope 8 Computing unit 9 Communication interface 0 Mobile device 1 Database 2 Display device 0 Test procedure Introduction Preparation Mixing Enrichment Evaluation

Claims

Patent claims 1. Sample container (10) for the detection of pathogens and / or cells (11) in a sample, in particular a blood sample, with a sample processing unit (12), comprising: - at least one receiving area (13) of the sample processing unit (12) for receiving the sample, - at least one processing area (14) of the sample processing unit (12) for processing the sample, - at least one mixing area (15) of the sample processing unit (12) for mixing the sample, wherein the mixing area (15) comprises a mixing structure (16), and - at least one enrichment region (17) for enriching the pathogens and / or cells (11) contained in the sample.

2. Sample container (10) according to claim 1, wherein the enrichment region (17) comprises at least one enrichment agent (19) and at least one dye (20), wherein the enrichment agent (19) comprises at least one antibody (21a) with at least one magnetic capture agent (22), in particular a magnetic microparticle, for capturing the pathogens and / or cells (11) in the sample, wherein the at least one dye (20) comprises at least one other antibody (21b) with at least one labeling agent (23), in particular a fluorescent labeling agent, for labeling the pathogens and / or cells (11) in the sample, wherein a sandwich structure (18) can preferably be produced by the at least one enrichment agent (19) and the at least one dye (20).

3. Sample container (10) according to one of the preceding claims, wherein the sample comprises a sample volume between 1 pL and 1 mL, preferably between 10 and 100 pL, particularly preferably between 20 and 30 pL.

4. Sample container (10) according to one of the preceding claims, wherein the enrichment region (17) is arranged close to the bottom within the sample container (10), wherein in particular at least one bottom (24) of the sample container (10) is transparent in optical quality.

5. Sample container (10) according to one of the preceding claims, wherein the processing area (14) comprises at least one lysis agent and / or at least one permeabilization agent for lysing and / or permeabilizing the pathogens and / or cells (11).

6. Test system (100) for detecting pathogens and / or cells (11) in a sample, in particular a blood sample, comprising: - at least one sample container (10) according to one of claims 1 to 5, wherein the sample container (10) comprises a sample comprising pathogens and / or cells (11), - at least one mixing unit (101) with a first sample holder (102) for receiving the sample container, wherein the mixing unit (101) is designed to mix at least the sample by means of movement, - at least one evaluation unit (103) with a second sample holder (104) for receiving the sample container, wherein the evaluation unit (103) is designed to enrich the pathogens and / or cells (11) in the enrichment region (17) of the sample container (10) and to visualize and detect at least one pathogen enriched in the enrichment region (17) and / or at least one cell (11) enriched in the enrichment region in the sample.

7. Test system (100) according to claim 6, wherein the sample container (10) in the processing area (14) has at least one lysis agent and / or at least one permeabilization agent for lysing and / or permeabilizing the cells and / or pathogens (11), wherein the mixing of the sample in the at least one mixing unit (101) comprises the lysis and / or permeabilization of the pathogens and / or cells (11).

8. Test system (100) according to claim 6 or 7, wherein the evaluation unit (103) comprises at least one magnet (105), wherein the evaluation unit (103) is designed to enrich at least one of the pathogens and / or at least one of the cells (11) by means of the magnet (105) in the enrichment region (17) of the sample container (10), wherein the at least one pathogen and / or the at least one cell (11) is bound to at least one enrichment agent (19), wherein the enrichment agent (19) comprises at least one antibody (21a) with a magnetic capture agent (22), in particular a magnetic microparticle, wherein the enrichment region (17) is arranged in particular close to the bottom within the sample container (10).

9. Test system (100) according to one of claims 6 to 8, wherein the evaluation unit (103) is designed to visualize and detect the pathogens and / or cells (11) through a bottom (24) of the sample container (11) in the enrichment region (17) of the sample container (11), wherein the bottom (24) is preferably transparent in optical quality, and / or wherein the evaluation unit (103) is designed to detect individual pathogens and / or individual cells (11) in the sample in the enrichment region (17) of the sample container (11).

10. Test system (100) according to one of claims 6 to 9, wherein the evaluation unit (103) comprises an image processing unit (106), wherein the image processing unit (106) is designed to visualize the pathogens and / or cells (11) individually, wherein the image processing unit (106) in particular comprises a, preferably inverse and / or automated, fluorescence microscope (107) which is designed to visualize the pathogens and / or cells (11) as fluorescent units.

11. Test system (100) according to one of claims 6 to 10, wherein the evaluation unit (103) comprises a computing unit (108) which is designed to quantify the at least one visualized pathogen and / or the at least one visualized cell (11) and / or to carry out pattern recognition.

12. Test system (100) according to one of claims 6 to 11, wherein the evaluation unit (103) comprises a communication interface (109) which is designed to transmit analysis data of the evaluation unit (103) to a mobile device (110) and / or a database (111), in particular a data cloud, and / or wherein the evaluation unit (103) comprises a display device (112), in particular a display, which is designed to display analysis data of the evaluation unit (103) to a user.

13. Test method (200) for detecting pathogens and / or cells (11) in a sample, in particular a blood sample, preferably using a test system (100) according to one of claims 6 to 12, comprising the following steps: Introducing (201) a sample, in particular a blood sample, into a receiving area (13) of a sample preparation unit (12) of a sample container (10), wherein the sample comprises pathogens and / or cells (11), - preparing (202) the sample in a preparation area (14) of the sample preparation unit (12) of the sample container (10), Mixing (203) the sample in a mixing area (15) of the sample preparation unit (12) of the sample container (10) by placing the sample container (10) into a mixing unit (101), - enriching (204) the pathogens and / or cells (11) in the sample in an enrichment area (17) of the sample container by placing the sample container in an evaluation unit, - evaluating (205) the enrichment area of the sample container by the evaluation unit, wherein the pathogens and / or cells in the sample are visualized and detected by means of the evaluation unit.

14. Test method (200) according to claim 13, wherein the processing (202) of the sample comprises lysis and / or permeabilization of the pathogens and / or cells (11), and / or wherein the enrichment (204) of the pathogens and / or cells (11) in the enrichment region (17) of the sample comprises magnetic capture and / or fluorescent labeling, and / or wherein the evaluation (205) of the enrichment region comprises quantification of the pathogens and / or cells (11) or pattern recognition, wherein the enrichment region (17) is arranged in particular close to the bottom within the sample container (10) and the evaluation (205) of the enrichment region (17) is carried out in particular through a bottom (24) of the sample container (10), wherein the bottom (24) is transparent and of optical quality.

15. Test method (200) according to claim 13 or 14, wherein the pathogens (11) comprise at least one malaria pathogen, in particular Plasmodium falciparum and / or Plasmodium vivax and / or Plasmodium ovale and / or Plasmodium knowlesi and / or Plasmodium malariae, and / or the cells (11) comprise at least one erythrocyte.

16. Test method (200) according to claim 13 or 14, wherein the pathogens (11) comprise at least one human immunodeficiency virus, and / or the cells (11) comprise at least one lymphocyte, preferably a T helper cell, particularly preferably a CD4 T cell and / or CD8 T cell.

17. Test method (200) according to one of claims 13 to 16 for diagnosing at least one disease, wherein in particular the disease is a disease caused by pathogens (11), in particular fungi, viruses and / or bacteria, and / or pathogens, in particular at least one from the group of Adenoviruses, Amylostoma, Ascaris, Babesia, Bacillus anthracis, Bordetella pertussis, Bordetella parapertussis, Borrelia recurrentis, Brucella sp., Campylobacter sp., Cestoda, Chlamydia psittaci, Clostridium botulinum, Corynebacterium diphtheriae, Coxiella burnetii, human pathogenic Cryptosporidium sp., Ebola virus, Echinococcus multilocularis, Echinococcus granulosus, Escherichia coli, enterohemorrhagic strains (EHEC), Eucestoda, Francisella tularensis, FSM E virus, yellow fever virus, Giardia lamblia, Haemophilus influenzae, hantaviruses, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus, hepatitis E virus, influenza viruses, Lassa virus, Legionella sp., human pathogenic Leptospira sp., Listeria monocytogenes, Marburgvirus, Measles virus, Mumps virus, Mycobacterium leprae, Mycobacterium tuberculosis / africanum, Mycobacterium bovis, Neisseria meningitidis, Norwalk-like virus, Poliovirus, Pseudomonas aeruginosa, Rabies virus, Rickettsia prowazekii, Rotavirus, Rubella virus, Salmonella Paratyphi, Salmonella Typhi, Schistosoma, Shigella sp., Taenia saginata, Taenia solium, Trichiuris, Trypanosoma, Trypanosoma brucei, Trypanosoma congolense, Trypanosoma vivax, Trichinella spiralis, Varicella-Zoster virus, Vibrio cholerae 1 and 139, Yersinia enterocolitica, Yersinia pestis, Treponema pallidum, HIV, Echinococcus sp., Plasmodium sp., Toxoplasma gondii, Streptococcus pneumoniae and / or Streptococcus aureus.

Images