Sample container, detection system and detection method for detecting pathogens and / or cells within 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, having 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 preparation area (14) of the sample processing unit (12) for preparing 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 area (17) for enriching pathogens and / or cells (11) contained in the sample. The invention also relates to a detection system (100) and a detection method (200) for detecting pathogens and / or cells (11) in a sample.

Description

Invention Field

[0001] This invention relates to sample containers for detecting pathogens and / or cells within a sample, as well as detection systems and corresponding detection methods for detecting pathogens and / or cells within a sample. Background Technology

[0002] Diagnostic testing systems for diseases are already publicly available in the technology. Here, not only laboratory-supported diagnostic systems, especially molecular diagnostic methods and immunological tests, are employed, but also so-called point-of-care testing (PoC testing).

[0003] Point-of-care (PoC) testing refers to patient-side diagnostics, which is not performed in a central laboratory but can be conducted directly at the bedside or in a testing station by a doctor or even a layperson. This provides the advantage that results are provided directly to the patient. PoC testing systems include, for example, pregnancy tests or coronavirus tests. In most cases, test strips are used for patient-side diagnostics, where a sample (e.g., blood or urine) is applied to the test strip and transported along the strip by capillary action. Here, the sample comes into contact with reagents, which cause a chemical reaction or binding in the presence of a target substance, thereby identifying a color change (e.g., a band-like appearance).

[0004] However, a drawback that has become apparent is that PoC assays in the form of test strips have low precision and sensitivity. Negative results may occur even in the presence of the target substance (such as a disease pathogen) at low concentrations.

[0005] This may be the case, for example, in cases of malignant malaria infection, where parasite levels may be very low, especially in pregnant women, asymptomatic carriers, and / or in the early stages of infection, so common detection systems are often insufficient to reliably detect the infection.

[0006] Another drawback of known Proof-of-Concept (PoC) tests is their inability to provide real-time measurement of pathogen status. In cases of malignant malaria infection, for example, only parasitic proteins that have circulated in the blood for several weeks are detected. This inability to perform real-time measurements limits possibilities such as monitoring drug efficacy.

[0007] Furthermore, test strips are inevitably exposed to the environment, so their results are particularly susceptible to the effects of high / low temperatures or air humidity, which can lead to distortion. Summary of the Invention

[0008] Therefore, the objective of this invention is to at least partially eliminate at least one of the aforementioned disadvantages. In particular, the objective of this invention is to provide a sample container, detection system, and detection method for detecting pathogens and / or cells within a sample, wherein the pathogens and / or cells can be detected accurately with high sensitivity in a low-cost and time-saving manner.

[0009] The above tasks are accomplished by a sample container having features of the first aspect of the invention, a detection system having features of the second aspect of the invention, and a detection method having features of the third aspect of the invention. Other features and details of the invention are derived from the dependent claims, the specification, and the drawings. Herein, the features and details described with respect to the sample container of the invention are obviously also applicable in relation to the detection system and / or the detection method of the invention, and vice versa; therefore, the disclosures regarding the various aspects of the invention are always cross-referenced or can be cross-referenced.

[0010] According to the present invention, according to a first aspect, a sample container for detecting pathogens and / or cells in a sample, particularly a blood sample, is provided, comprising a sample processing unit including: - At least one receiving area of ​​the sample processing unit for receiving samples. - At least one preparation area for preparing samples in this sample processing unit. - At least one mixing region for mixing samples in the sample processing unit, wherein the mixing region includes a mixing structure, and - At least one enrichment zone for enriching pathogens and / or cells contained in the sample.

[0011] The sample container may specifically refer to a reaction vessel. The sample container is preferably made at least partially of polypropylene and / or a cyclic olefin copolymer (ethylene-norbornene copolymer). The sample container may particularly be conical and / or produced by injection molding. Additionally, the sample container may have multiple walls, wherein the angle of the sample container walls preferably tapers gradually towards the bottom. It may also be specified that the sample container has a flat bottom. It is also conceivable that the sample container has a circular bottom surface, particularly having a diameter between 0.1 µm and 10 mm, preferably between 0.5 µm and 1 mm, and particularly preferably between 1 µm and 100 µm.

[0012] The pathogen may preferably include at least one Plasmodium and / or at least one human immunodeficiency virus (HIV). Additionally, the cells may include at least one erythrocyte and / or lymphocyte, particularly helper T cells.

[0013] The sample may be, in particular, a blood sample, wherein the sample preferably includes capillary blood and / or venous whole blood from a fingertip. In addition, the sample may include other bodily fluids, such as urine, saliva, synovial fluid, cerebrospinal fluid, plasma, serum, tears, sweat, lymph, and / or interstitial fluid.

[0014] The sample container may have a sample processing unit, wherein the sample processing unit may include a receiving area, a preparation area and / or a mixing area.

[0015] The receiving area can refer to the region of a sample container where samples, especially blood samples, are added. Adding samples to the sample container can preferably be done manually or automatically.

[0016] The preparation zone can refer to a sample container area where pathogens and / or cells contained in the sample are pretreated. The preparation zone preferably includes reagents, particularly lyophilized and / or dried reagents. The reagents may particularly include at least one lysing agent and / or at least one permeabilizing agent and / or at least one enriching agent and / or at least one staining agent and / or at least one stabilizer. Pathogens and / or cells are pretreated, particularly by contacting the pathogens and / or cells with the preferably dissolved reagent.

[0017] Lysing agents can specifically refer to formulations used to almost completely disrupt the outer and / or inner cell membranes of pathogens and / or cells. Permeabilizing agents can refer to formulations used to create temporary and / or persistent permeability to the outer and / or inner cell membranes of pathogens and / or cells. Enriching agents can preferably comprise at least one antibody having a magnetic trapping agent for capturing pathogens and / or cells from a sample. Staining agents can preferably contain at least one antibody having a labeling agent for labeling pathogens and / or cells within a sample. Stabilizers can refer to substances that maintain the stability and functionality of pathogens and / or cells and / or enriching agents and / or staining agents.

[0018] The mixing zone can refer to the sample container area where sample mixing takes place. For this purpose, a mixing structure can be provided within the mixing zone of the sample container. The mixing structure can particularly have at least one mixing element, preferably in the form of a bead. Alternatively, the mixing structure can have at least one mixing element in the form of a protrusion, particularly in the form of a wing and / or edge and / or mound-like protrusion arranged on the inner wall of the sample container within the mixing zone. Sample mixing is preferably carried out by centrifugal and / or eccentric motion. The mixing element places the liquid to be mixed in a more intense motion. During mixing, centrifugal force first presses the sample against the edge of the sample container. The mixing element prevents the sample from completely adhering to the edge of the sample container and thus promotes more effective dispersion of the sample and / or reagent components. They intentionally introduce the mixture, especially the liquid, into the mixing element area, which helps the sample and / or reagent to mix more uniformly in the mixture. The mixing element generates a more intense vortex in the liquid flow. Optimizing the flow behavior of the mixture, especially the liquid, through the mixing element can further improve the mixing of the sample and / or reagent and make it more efficient.

[0019] Additionally, the sample container has an enrichment zone. The enrichment zone can refer to a region of the sample container where pathogens and / or cells contained in the sample are concentrated. This enrichment zone preferably has at least one enriching agent and / or at least one staining agent. The enriching agent may, in particular, have at least one antibody for enriching pathogens and / or cells contained in the sample, wherein the at least one antibody includes a magnetic trapping agent, particularly magnetic microparticles, for capturing pathogens and / or cells within the sample. The staining agent may include at least another antibody having a labeling agent, particularly a fluorescent labeling agent, for labeling pathogens and / or cells within the sample. Preferably, a sandwich structure can be created using the enriching agent, the staining agent, and a single pathogen and / or a single cell.

[0020] Enrichment within the enrichment zone can preferably be performed using a magnet, particularly at the bottom of the sample container, by means of magnetic force. The at least one enriching agent and / or at least one staining agent can be arranged within the sample container, particularly in lyophilized and / or dried reagent form, wherein the at least one enriching agent and / or at least one staining agent is preferably pre-presented on the inner wall and / or bottom of the sample container. Additionally, the reagents may particularly include at least one lysing agent and / or permeabilizing agent and / or at least one stabilizer. Furthermore, pre-preservation saves additional time because no reagent addition is required, allowing pre-fabricated batch sample containers to be used outside the laboratory at the patient's side.

[0021] The receiving area, preparation area, mixing area, and enrichment area of ​​the sample container can preferably exist in the form of homogeneous regions with continuous, gradual transitions. Furthermore, the receiving area, preparation area, mixing area, and enrichment area of ​​the sample container can exist as a single region, wherein these regions preferably completely overlap.

[0022] Therefore, it enables highly accurate and sensitive detection of cells and / or pathogens in samples, especially blood samples. This mixing structure allows for particularly efficient sample mixing, thereby optimizing the enrichment of pathogens and / or cells within the sample, and consequently improving detection sensitivity. It also allows for the detection of low concentrations of pathogens and / or cells within the sample.

[0023] Within the scope of this invention, the enrichment region may further be specified to include at least one enriching agent and at least one staining agent, wherein the enriching agent includes at least one antibody having at least one magnetic trapping agent, particularly magnetic microparticles, for capturing pathogens and / or cells within the sample, wherein the at least one staining agent includes at least one other antibody having at least one labeling agent, particularly fluorescent labeling agent, for labeling pathogens and / or cells within the sample, wherein, preferably, a sandwich structure can be generated by the at least one enriching agent and the at least one staining agent.

[0024] In other words, the enrichment region may contain at least one enriching agent and / or at least one staining agent. The enriching agent particularly includes at least one antibody carrying a magnetic trapping agent for capturing pathogens and / or cells from the sample. The staining agent preferably includes at least one other antibody carrying a labeling agent for labeling pathogens and / or cells within the sample. A sandwich structure can preferably be created by using at least one enriching agent, at least one staining agent, and at least one pathogen and / or cell.

[0025] The antibody is preferably constructed as a monoclonal antibody, capable of specifically binding to target structures on the surface of pathogens and / or cells. The target structures may, in particular, be epitopes on pathogens and / or cell surface antigens.

[0026] It may be specified that at least one antibody is attached to a magnetic trapping agent, and at least another antibody is attached to a labeling agent.

[0027] Magnetic trapping agents may particularly include magnetic microparticles, preferably Dynabead and / or Turbobead. Here, the magnetic microparticles may have a diameter from 0.05µm to 20µm, preferably from 0.07µm to 10µm, and particularly preferably from 0.5µm to 5µm.

[0028] The labeling agent may include fluorescent labeling agents, especially fluorescent dyes. Fluorescent labeling agents are preferably excited by optical excitation to emit a fluorescent signal, which can be detected by means of a fluorescence microscope. Optical excitation is particularly performed in the near-infrared range, preferably between 700 and 2000 nm, and especially at wavelengths between 700 and 900 nm.

[0029] Antibodies linked to magnetic trapping agents and antibodies linked to labeling agents can bind to different epitopes on pathogens and / or cell surface antigens, respectively. This allows for the formation of a sandwich structure produced by two antibodies and a single pathogen and / or a single cell.

[0030] This provides the following advantages: by applying a magnet to the enrichment area of ​​the sample container, at least one antibody linked to at least one magnetic trapping agent can be enriched by magnetic force, wherein the at least one antibody also binds to target structures on the surface of pathogens and / or cells contained in the sample. Therefore, pathogens and / or cells contained in the sample can be enriched within the enrichment area of ​​the sample container, and even low concentrations of pathogens and / or cells in the sample can be detected.

[0031] Since pathogens and / or cells also bind to at least one antibody labeled with a marker, enriched pathogens and / or cells can be detected very accurately, 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 and cost-effectively. When optically excited in the wavelength range of 700 to 2000 nm, the autofluorescence of biological materials is particularly low, thus allowing for better signal quality under fluorescently labeled optical excitation to detect pathogens and / or cells contained in the sample.

[0032] Within the scope of this invention, it is conceivable that the sample includes a sample volume between 1 µL and 1 mL, preferably between 10 and 100 µL, and particularly preferably between 20 and 30 µL.

[0033] Therefore, even with a large sample volume, it can detect pathogens and / or cells in the sample with high sensitivity. In particular, it can also achieve the detection of low concentrations of pathogens and / or cells with a large sample volume through enrichment.

[0034] It is also conceivable that the enrichment region is arranged near the bottom inside the sample container, wherein, in particular, at least the bottom of the sample container is constructed with optical-grade transparency.

[0035] "Near the bottom" can specifically mean that the enrichment region preferably extends between 0 µm and 200 µm above the bottom of the sample container, more preferably between 0 µm and 50 µm, and especially preferably between 0 µm and 25 µm. 0 µm means that the enrichment region starts directly from the bottom.

[0036] The bottom of the sample container is preferably planar. Alternatively, the bottom of the sample container can be made of a cyclic olefin copolymer (ethylene norbornene copolymer), glass, polyethylene terephthalate, polymethyl methacrylate, and / or polycarbonate. Here, the bottom of the sample container is preferably made of a cyclic olefin copolymer (ethylene norbornene copolymer). Cyclic olefin copolymers have the advantage of excellent optical properties, such as high light transmittance and low autofluorescence.

[0037] "The bottom of the sample container is constructed with optical-grade transparency" specifically refers to its high light transmittance. This allows sufficient fluorescence to penetrate the bottom of the sample container and enter the sample during fluorescence excitation, making the pathogens and / or cells contained in the sample visible. Furthermore, because the bottom of the sample container is constructed with optical-grade transparency, sufficient fluorescence from the sample can be detected by sensors located at the bottom of the sample container and / or perceived by the user's eye during fluorescence excitation.

[0038] Furthermore, the sample container can be constructed with complete optical-grade transparency, preferably made of a cyclic olefin copolymer (ethylene norbornene copolymer). This ensures that when fluorescence is incident laterally into the sample container, sufficient fluorescence from the sample can be detected by the sensor located at the bottom of the sample container and / or perceived by the user's eye under fluorescence excitation.

[0039] The sensor is preferably designed as a photomultiplier tube (PMT) or a charge-coupled device (CCD).

[0040] Here, thanks to the optical quality at the bottom, exceptionally uniform image brightness, high image contrast, and / or high resolution can be achieved when creating images by the evaluation unit. Furthermore, this minimizes geometric distortions and optical aberrations, such as spherical or chromatic aberration, coma, and astigmatism. Preferably, pathogens and / or cells contained in the sample can be identified on the image created by the evaluation unit.

[0041] This provides the following overall advantages: precise and time-saving detection of pathogens and / or cells within the sample can be achieved through an optically advantageous bottom construction, and errors that may be caused by optical interference during visualization can be avoided.

[0042] It is also conceivable that the preparation zone has at least one lysis agent and / or at least one permeabilizing agent for the lysis and / or permeabilization of pathogens and / or cells.

[0043] The lysing agent may, for example, contain Triton X100 and / or deoxycholate. The permeabilizing agent preferably includes saponins and / or ammonium chloride. "Lystagnation" may specifically refer to the near-complete disruption of the outer and / or inner cell membranes of pathogens and / or cells. "Permeabilization" may specifically refer to the process of creating temporary and / or permanent permeability to the outer and / or inner cell membranes of pathogens and / or cells. Lysis and / or permeabilization, respectively, allow access to the internal structures of pathogens and / or cells.

[0044] The advantage of lysing and / or permeating the cells contained in a sample is that it also allows for the detection of intracellular structures. For example, disease pathogens (such as parasites, bacteria, or viruses) may enter so-called host cells, making the exposure of intracellular structures crucial for the detection of these pathogens. Additionally, lysis and / or permeation can expose the inner surface of the cell membrane, allowing for the detection of structures arranged there, such as specific proteins.

[0045] When pathogens and / or cells contained in a sample are lysed and / or permeated, biomolecular components of the pathogens and / or cells (such as proteins, nucleic acids, or lipids) can be released for highly specific detection.

[0046] Therefore, it is possible to achieve very accurate detection of pathogens and / or cells in general, because specific components of pathogens and / or cells can be detected.

[0047] The aforementioned task is also accomplished through a second aspect of the invention. The second aspect of the invention includes a detection system for detecting pathogens and / or cells in a sample, particularly a blood sample, comprising: - At least one sample container according to a first aspect of the invention, wherein the sample container contains a sample comprising pathogens and / or cells. - At least one mixing unit having a first sample holder for receiving a sample container, wherein the mixing unit is designed to mix at least the sample by means of movement. - At least one evaluation unit having a second sample holder for receiving a sample container. The evaluation unit is designed to enrich pathogens and / or cells within the enrichment zone of the sample container, and to visualize and detect at least one pathogen and / or at least one cell enriched in the enrichment zone within the sample.

[0048] The sample container preferably also contains, in particular, pre-stored reagents. The reagents may preferably be formed in a lyophilized and / or dried form. The reagents particularly include at least one lysing agent and / or at least one permeabilizing agent and / or enriching agent and / or at least one staining agent and / or at least one stabilizer. The at least one lysing agent may preferably include Triton X, for almost complete disruption of the cell membranes of pathogens and / or cells. The at least one permeabilizing agent may particularly include saponins, for generating temporary and / or persistent permeability to pathogens and / or cells. The at least one enriching agent may preferably include at least one antibody having a magnetic trapping agent for capturing and enriching pathogens and / or cells contained in the sample. The at least one staining agent may preferably have at least one antibody having a labeling agent for labeling pathogens and / or cells contained in the sample. The at least one stabilizer may preferably have a polysaccharide for maintaining the components of the sample and / or reagents.

[0049] The mixing unit may, in particular, have an eccentric mixer. Additionally, the mixing unit preferably includes a rotary mixer and / or a vibratory mixer.

[0050] An eccentric mixer can refer to a mechanical device that mixes components of a sample and / or reagent by eccentric rotation.

[0051] Mixing of samples and / or reagents within a sample container is particularly preferably performed using an eccentric mixer via elliptical and / or rotational motion, wherein the sample container also rotates itself. Alternatively or additionally, mixing of samples within a sample container can be performed by alternating vibrations at the center of the sample container.

[0052] Additionally, the mixing of samples and / or reagents within the sample container, particularly using an eccentric mixer, may include sample activation and / or sample lysis and / or permeabilization. Sample activation may, in particular, refer to the generation of a sandwich complex consisting of an enriching agent and a staining agent, as well as pathogens and / or cells. The enriching agent preferably comprises at least one antibody having a magnetic trapping agent, particularly magnetic microparticles, for capturing pathogens and / or cells within the sample. The staining agent particularly comprises at least another antibody having a labeling agent, particularly a fluorescent labeling agent, for labeling pathogens and / or cells within the sample. The at least two antibodies may bind, in particular, to epitopes on pathogen and / or cell surface antigens, preferably different epitopes, thus creating a sandwich structure.

[0053] The mixing unit may include a first sample holder, preferably in the form of a clamping device and / or fixture. Alternatively or additionally, the first sample holder may also be designed as a recess within the mixing unit, into which a sample container can be inserted and held in place during mixing, particularly by means of a locating pin and / or by pressing. Mixing of samples within the mixing unit can be particularly automated.

[0054] Furthermore, the mixing unit can be designed to regulate the temperature of the sample within the sample container. Specifically, the temperature is specified to be between 0 and 100°C, preferably between 20 and 60°C, and particularly preferably between 30 and 40°C.

[0055] The evaluation unit may specifically refer to a device designed to enrich and / or visualize and detect pathogens and / or cells contained in a sample. For this purpose, the evaluation unit may preferably include a magnet for magnetically enriching the pathogens and / or cells contained in the sample, wherein, in particular, the pathogens and / or cells are each bound to at least one enriching agent, wherein the enriching agent comprises at least one antibody with a magnetic trapping agent, particularly with magnetic particles.

[0056] Additionally, the evaluation unit may include an image processing unit, particularly equipped with a microscope, preferably a fluorescence microscope, for visualizing and detecting pathogens and / or cells contained in the sample. For this purpose, the pathogens and / or cells are preferably each bound to at least one staining agent, wherein the staining agent comprises at least one antibody with a labeling agent, particularly a fluorescent labeling agent. Visualizing and detecting pathogens and / or cells contained in the sample via the evaluation unit can preferably be performed by using fluorescence to excite the pathogens and / or cells with fluorescent labels.

[0057] The evaluation unit may include a second sample holder, preferably in the form of a clamping device and / or fixture. Alternatively or additionally, the second sample holder may also be designed as a recess within the evaluation unit, wherein the sample container can be inserted into the recess and held in place during mixing, particularly by means of locating pins and / or by pressing. Visualization and detection of pathogens and / or cells contained in the sample can preferably be automated.

[0058] This provides the following overall advantages: it enables particularly time-saving and low-cost detection of pathogens and / or cells within a sample, where even low concentrations of pathogens and / or cells can be accurately detected.

[0059] Within the scope of this invention, it may also be specified that the sample container has at least one lysis agent and / or at least one permeabilizing agent in the preparation area for the lysis and / or permeabilization of cells and / or pathogens, wherein the mixing of the sample in at least one mixing unit includes the lysis and / or permeabilization of pathogens and / or cells.

[0060] The pyrolysis agent may include, for example, Triton X100 and / or deoxycholate. The permeabilizing agent preferably includes saponins and / or ammonium chloride.

[0061] This provides the following advantages: it enables particularly time-saving and accurate detection of pathogens and / or cells within a sample. Through the lysis and / or permeation of pathogens and / or cells, the specific biomolecular structures of the pathogens and / or cells can be released, thereby ensuring accurate detection.

[0062] Within the scope of this invention, the evaluation unit may also include at least one magnet, wherein the evaluation unit is designed to enrich at least one pathogen and / or at least one cell in an enrichment area of ​​the sample container by means of the magnet, wherein the at least one pathogen and / or at least one cell binds to at least one antibody having a magnetic trapping agent, particularly having magnetic particles, wherein the enrichment area is arranged particularly close to the bottom within the sample container.

[0063] The at least one magnet may be, in particular, a toroidal magnet and / or a cylindrical magnet. The at least one magnet may preferably be designed as a neodymium magnet and / or an on / off electromagnet and / or an array of permanent magnets. It may also be specified that the at least one magnet is arranged, particularly within the evaluation unit, below the second sample holder. This preferably achieves enrichment in an enrichment region near the bottom of the sample container. "Near the bottom" can mean that the enrichment region extends preferably between 0 µm and 200 µm, more preferably between 0 µm and 50 µm, and particularly preferably between 0 µm and 25 µm above the bottom of the sample container. 0 µm means that the enrichment region begins directly from the bottom.

[0064] It can also be specified that the enrichment of pathogens and / or cells contained in the sample is carried out using antibodies linked to magnetic trapping agents, particularly those linked to magnetic microparticles. These antibodies are able to specifically bind to target structures on the surface of pathogens and / or cells. By inserting the sample container into the second sample scaffold of the evaluation unit, a magnet can use magnetic force to enrich pathogens and / or cells with the magnetic trapping agent within the enrichment area of ​​the sample container.

[0065] This provides the following advantages: it enables the detection of pathogens and / or cells enriched in the enrichment area of ​​the sample container with high sensitivity.

[0066] Within the scope of this invention, it is also conceivable that the evaluation unit is designed to penetrate the bottom of the sample container to visualize and detect pathogens and / or cells within the enrichment area of ​​the sample container, wherein the bottom is preferably constructed with optical-grade transparency, and / or wherein the evaluation unit is designed to detect individual pathogens and / or individual cells within the sample in the enrichment area of ​​the sample container.

[0067] "The bottom of the sample container is constructed with optical-grade transparency" specifically refers to a bottom with high light transmittance. This allows sufficient fluorescence to enter the sample when it passes through the bottom of the sample container, enabling the pathogens and / or cells contained in the sample to be visible upon fluorescence excitation. Furthermore, because the bottom of the sample container is constructed with optical-grade transparency, sufficient fluorescence from the sample can be detected by sensors located at the bottom of the sample container and / or perceived by the user's eye upon fluorescence excitation.

[0068] Furthermore, the sample container can be constructed with near-complete optical-grade transparency, preferably made of a cyclic olefin copolymer (ethylene norbornene copolymer). This allows sufficient fluorescence from the sample to be detected by sensors located at the bottom of the sample container and / or perceived by the user's eye when lateral fluorescence is incident into the sample container under fluorescence excitation.

[0069] The sensor can preferably be designed as a photomultiplier tube (PMT) or a charge-coupled device (CCD).

[0070] Due to the optical quality at the bottom, exceptionally uniform image brightness, high image contrast, and / or high resolution can be achieved. Furthermore, this minimizes geometric distortions and optical aberrations such as spherical or chromatic aberration, coma, and astigmatism.

[0071] Visualization and detection of individual pathogens and / or individual cells within the enrichment zone of the sample container can be achieved using at least one staining agent. This at least one staining agent preferably comprises at least one antibody linked to a labeling agent, particularly a fluorescent labeling agent. The at least one antibody specifically binds to an epitope on the surface antigens of the pathogens and / or cells contained in the sample. Pathogens and / or cells contained in the sample can be enriched within the enrichment zone of the sample container by using at least one enriching agent comprising at least one other antibody linked to a magnetic trapping agent, particularly a magnetic microparticle. The at least one antibody preferably binds to another epitope on the surface antigens of the pathogens and / or cells, particularly a different epitope. Pathogens and / or cells are enriched within the enrichment zone, particularly by placing a magnet within the enrichment zone, especially within the evaluation unit.

[0072] By using a staining agent containing a labeling agent, preferably a fluorescent labeling agent, visualization and detection of individual enriched pathogens and / or cells can be achieved, particularly by optical excitation and with the aid of a microscope, preferably a fluorescence microscope.

[0073] Because pathogens and / or cells are enriched within the enrichment area of ​​the sample container and can subsequently be visualized and detected, pathogens and / or cells within the sample can be detected with high sensitivity. Since individual pathogens and / or cells can be detected, even low concentrations of pathogens and / or cells within the sample can be detected. Therefore, disease transmission can be prevented, as even asymptomatic carriers and / or pregnant women with very low pathogen and / or cell loads can be identified and treated. Furthermore, drug monitoring can be performed by detecting individual pathogens and / or cells, as changes in pathogen and / or cell numbers can be detected very precisely.

[0074] It is also conceivable that the evaluation unit includes an image processing unit, wherein the image processing unit is designed to visualize pathogens and / or cells individually, wherein the image processing unit particularly includes, in particular, a fluorescence microscope preferably inverted and / or automated, designed to visualize pathogens and / or cells as fluorescence units.

[0075] The image processing unit may preferably include hardware and / or software, wherein the image processing unit is particularly designed to analyze, process and interpret image data of the enriched region of the sample container.

[0076] The image processing unit may preferably include, in particular, an inverted and / or automated fluorescence microscope, wherein the fluorescence microscope may have an optical magnification unit.

[0077] When using a fluorescence microscope, pathogens and / or cells bind to a staining agent containing at least one antibody and having at least one fluorescent labeling agent attached to it; they can be detected by fluorescence excitation after being enriched in an enrichment area of ​​a sample container.

[0078] The image processing unit, preferably a fluorescence microscope, may include an objective lens and / or a light source. The objective lens is preferably positioned below the sample container, while the light source is preferably positioned above or to the side of the sample container, and / or below it. The light source can produce radiation, preferably in the near-infrared range and particularly including wavelengths between 700 and 2000 nm, preferably between 700 and 900 nm.

[0079] The image processing unit, preferably a fluorescence microscope, is particularly designed to generate a series of images (a Z-axis stacked sequence) created by focusing on different layers of the sample within the sample container. Each image in the Z-axis stacked sequence displays a specific cross-section of the sample along the Z-axis (vertical axis). Here, the image processing unit can be designed to create 10 to 50 images, preferably 15 to 40, and particularly preferably 20 to 30 images at different layers of the sample. Alternatively, the image processing unit can be designed to create a single image using the maximum intensity projection of the Z-axis stacked sequence. Pathogens and / or cells contained in the sample are preferably visible on this image.

[0080] This offers the advantages of enabling highly accurate detection of pathogens and / or cells within a sample in a low-cost and time-saving manner. Furthermore, fluorescence detection offers advantages over common detectors (such as those using Giemsa staining) in terms of higher sensitivity and specificity. Additionally, waiting times can be reduced because only a short incubation period needs to be considered.

[0081] Within the scope of this invention, the evaluation unit may also include a computing unit designed to quantify the at least one visualized pathogen and / or the at least one visualized cell, and / or perform pattern recognition.

[0082] The computing unit can be designed to be mobile or fixedly connected to the evaluation unit.

[0083] The computing unit preferably includes self-learning software. The computing unit may also be designed to determine the number of visualized pathogens and / or cells. Additionally or alternatively, the computing unit may be designed to perform pattern recognition, wherein, in particular, the following steps are performed: - Perform maximum intensity projection on the Z-axis stacked sequence to produce a single image of the sample container enrichment region, wherein the Z-axis stacked sequence is created by the image processing unit of the evaluation unit. - Adjust, especially enhance, the image contrast. - Perform background subtraction on the image. - Identify patterns of pathogens and / or cells visualized in the image, wherein the image contains, in particular, 1 to 20 megapixels, preferably 1 to 10 megapixels, and especially preferably 1 to 5 megapixels.

[0084] Additionally, size-based filtering is performed in pattern recognition, where objects visible in the image that exceed a preset size range (preferably larger than or smaller than 4 pixels) and / or whose shape exceeds a preset range (preferably 2×2 pixels) can be filtered out. Pathogens and / or cells contained in the sample are preferably considered as objects visible in the image.

[0085] This provides the following advantages: ensuring highly sensitive and accurate detection of pathogens and / or cells within a sample. By accurately determining the number of pathogens and / or cells in the sample, it is possible to determine disease progression, identify and treat asymptomatic carriers with low pathogen and / or cell loads, and identify drug resistance. Pattern recognition offers the particularly time-saving and cost-effective advantage of detecting pathogens and / or cells within a sample.

[0086] Within the scope of this invention, it is also conceivable that the evaluation unit includes a communication interface designed to transmit the analysis data of the evaluation unit to a mobile device and / or a database, particularly a data cloud, and / or wherein the evaluation unit includes a display device, particularly a display, designed to display the analysis data of the evaluation unit to a user.

[0087] The analytical data may preferably include the number of pathogens and / or cells and / or pattern recognition data within the sample. Additionally, the analytical data may include personnel-related data, preferably basic data and / or medical data. Basic data may in particular include the person's name, date of birth, and / or gender. Medical data may in particular include the person's medical history and / or physician's reports.

[0088] It can also be specified that the analyzed data includes information about pathogen and / or cell testing for a particular individual, preferably allowing for a comparison of pathogen and / or cell counts within the sample after retesting. This allows, for example, the early identification of drug resistance.

[0089] The communication interface can be designed, in particular, to transmit analytical data in a wired and / or wireless manner, preferably via Bluetooth and / or WLAN (Wireless Local Area Network).

[0090] Mobile devices can preferably be tablets, smartphones, or smartwatches.

[0091] The database can preferably include a cloud-based data platform, through which analytical data can be accessed. This data includes, in particular, anonymized information about pathogens and / or cells within the tested samples (positive test results), especially in conjunction with the location of the test. This allows for the creation of prevalence maps, which can, in particular, provide access to medical personnel and medications for locations with high detection rates.

[0092] This offers the following advantages: the spread of disease can be located and contained using databases. Furthermore, patients or doctors can read results directly on mobile devices or displays, saving time as complex laboratory reports need not be analyzed.

[0093] The aforementioned task is also accomplished through a third aspect of the invention. The third aspect of the invention includes a method for detecting pathogens and / or cells in a sample, particularly a blood sample, preferably using a detection system according to the second aspect of the invention, comprising the following steps: - The receiving area of ​​a sample processing unit that introduces samples, particularly blood samples, into a sample container, wherein the sample contains pathogens and / or cells. - Prepare the sample in the preparation area of ​​the sample processing unit of the sample container. - By feeding the sample container into the mixing unit, the sample is mixed in the mixing area of ​​the sample processing unit within the sample container. - By feeding the sample container into the evaluation unit, pathogens and / or cells within the sample are enriched in the enrichment zone of the sample container. - The enrichment region of the sample container is evaluated by an evaluation unit, which is used to visualize and detect pathogens and / or cells within the sample.

[0094] The testing method can preferably be performed by medical personnel, laypeople (persons without medical training), or patients. Furthermore, the testing method can be performed automatically or manually.

[0095] Furthermore, the assessment of enriched regions within a sample container can include real-time quantification of pathogens and / or cells within the sample. The assessment of enriched regions within a sample container may also include visualization and detection of individual pathogens and / or cells.

[0096] In addition, reagents, particularly lyophilized and / or dried reagents, can be pre-stored in the sample container. These reagents may include at least one lysing agent and / or at least one permeabilizing agent and / or at least one enriching agent and / or at least one staining agent and / or at least one stabilizer. The lysing agent may particularly contain Triton X100 and / or deoxycholate. The permeabilizing agent preferably includes saponins and / or ammonium chloride. The enriching agent may preferably contain at least one antibody capable of binding to pathogens and / or cells within the sample, wherein said at least one antibody may contain a magnetic trapping agent, preferably magnetic microparticles. The staining agent preferably includes another antibody with a labeling agent, preferably a fluorescent labeling agent, wherein the labeling agent is particularly capable of labeling pathogens and / or cells within the sample. The two antibodies and a single pathogen and / or a single cell may preferably form a sandwich structure. The stabilizer may particularly include polysaccharides, preferably dextran.

[0097] It may be specified to add the sample to be tested, especially a blood sample, to a reagent located within a sample container, particularly a lyophilized and / or dried reagent. It is also conceivable to expose samples containing pathogens and / or cells to the reagent.

[0098] Subsequently, the sample container, containing, in particular, lyophilized and / or dried reagents and samples, can preferably be fed into a first sample holder of a mixing unit, particularly equipped with an eccentric mixer. The mixing unit preferably allows for sample mixing, wherein, during mixing, in particular, lysis and / or permeation of pathogens and / or cells within the sample, and / or enrichment of pathogens and / or cells within the sample in an enrichment zone of the sample container, and / or labeling of pathogens and / or cells within the sample can be performed.

[0099] It is also conceivable that the sample container is subsequently placed into a second sample holder in the evaluation unit. The evaluation unit may preferably include a magnet and / or a microscope, particularly a fluorescence microscope. Pathogens and / or cells are preferably bound to at least one enriching agent having at least one antibody, which may be linked to a magnetic trapping agent. Pathogens and / or cells within the sample can thus be enriched in the enrichment area by magnetic force using a magnet. Additionally, pathogens and / or cells are particularly bound to at least one staining agent and can be visualized and detected using a microscope, the staining agent having at least another antibody, which may be linked to a labeling agent.

[0100] This provides the advantage of enabling accurate detection of pathogens and / or cells within a sample in a simple, low-cost, and time-saving manner. Furthermore, the sensitivity of this detection method can be improved, particularly through the enrichment of pathogens and / or cells.

[0101] Within the scope of this invention, sample preparation may also include lysis and / or permeation of pathogens and / or cells, and / or wherein enrichment of pathogens and / or cells within an enrichment zone of the sample container includes magnetic capture and / or fluorescent labeling, and / or wherein evaluation of the enrichment zone includes quantification or pattern recognition of pathogens and / or cells, wherein the enrichment zone is disposed particularly close to the bottom within the sample container, and the evaluation of the enrichment zone is particularly performed through the bottom of the sample container, wherein the bottom is constructed with optically grade transparency.

[0102] The lysis agent may, for example, contain Triton X100 and / or deoxycholate. The permeabilizing agent preferably contains saponins and / or ammonium chloride. Lysis and / or permeabilization respectively allow access to the internal structures of pathogens and / or cells.

[0103] To enrich pathogens and / or cells contained in a sample within an enrichment zone, the pathogens and / or cells may bind to at least one enriching agent. The at least one enriching agent preferably comprises at least one antibody with a magnetic trapping agent, preferably magnetic microparticles. By applying a magnet, pathogens and / or cells with the magnetic trapping agent can be concentrated within the enrichment zone of the sample container.

[0104] The pathogens and / or cells contained in the sample are preferably also bindable to at least one staining agent. The at least one staining agent preferably contains another antibody labeled, preferably a fluorescent labeling agent.

[0105] The at least two antibodies can bind to different epitopes on pathogens and / or cell surface antigens, respectively. Therefore, a sandwich structure can be generated using the at least one enriching agent, the at least one staining agent, and a single pathogen and / or a single cell. The sandwich structure is preferably enriched, particularly by means of a magnet, within the enrichment area of ​​the sample container.

[0106] The enriched pathogens and / or cells are preferably visualized and detected individually, especially by fluorescence microscopy.

[0107] Overall, the following advantages can be obtained: the sensitivity and specificity of in-sample pathogen and / or cell detection are further improved because the biomolecular structure of pathogens and / or cells is also detected, and even low concentrations of pathogens and / or cells can be detected through enrichment.

[0108] Within the scope of this invention, it is also conceivable that the pathogen includes at least one malaria pathogen, particularly Plasmodium falciparum and / or Plasmodium vivax and / or Plasmodium ovale and / or Plasmodium norovirus and / or Plasmodium malariae, and / or the cell includes at least one red blood cell.

[0109] This yields the following advantages: the detection method can be used to detect malaria. Malaria is a deadly disease that spreads precisely in areas lacking laboratory diagnostic capabilities. Therefore, the ability for even laypeople (those without medical training) and / or patients themselves to perform low-cost and time-saving malaria pathogen testing is crucial for malaria control.

[0110] It is also conceivable that the pathogen includes at least one HIV virus, and / or the cells include at least one lymphocyte, preferably a helper T cell, especially preferably a CD4-T cell and / or a CD8-T cell.

[0111] It can be specified that, when the enrichment area of ​​the sample container is evaluated by the evaluation unit, a reduction in the number of helper T cells, particularly CD4-T cells and / or CD8-T cells, can be detected. It is conceivable that the enrichment of helper T cells is performed using at least one enriching agent comprising at least one antibody, which is attached to at least one magnetic trapping agent, particularly at least one magnetic microparticle. Here, the at least one antibody with the magnetic trapping agent can bind to epitopes on the surface antigens of helper T cells. Detection of the enriched helper T cells can be performed using a staining agent, wherein the staining agent preferably comprises another antibody, which is attached to at least one labeling agent, particularly at least one fluorescent labeling agent. Here, the at least one other antibody with the labeling agent binds to a different epitope on the surface antigens of helper T cells. This can thus produce a sandwich complex comprising the two antibodies and helper T cells, wherein the sandwich complex can be enriched in the enrichment area of ​​the sample container by means of a magnet and visualized and detected by an image processing unit, particularly equipped with a fluorescence microscope.

[0112] This provides the following advantages: by using this detection method to detect helper T cells in a sample with high sensitivity, and especially to monitor the number of helper T cells in the sample, it is possible to identify the onset of AIDS caused by HIV virus that invades helper T cells.

[0113] Within the scope of this invention, diseases can also be defined as those caused by pathogens, particularly fungi, viruses, and / or bacteria and / or pathogenic agents, especially at least one of the following groups: adenovirus, hookworms, ascarids, babesi, Bacillus anthracis, Bordetella pertussis, Bordetella parapertussis, Treponema relapsingis, Brucella, Campylobacter, tapeworms, Chlamydia psittaci, Clostridium botulinum, Corynebacterium diphtheriae, Coxsella bengali, Cryptosporidium spp., Ebola virus, Echinococcus multilocularis, Echinococcus granulosus, Escherichia coli, enterohemorrhagic Escherichia coli (EHEC), tapeworms, Tulafrancella, tick-borne encephalitis virus, yellow fever virus, Giardia lamblia, Haemophilus influenzae, Hantavirus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus. Hepatitis E virus, influenza virus, Lassa virus, Legionella spp., human pathogenic Leptospira spp., Listeria monocytogenes, Marburg virus, measles virus, mumps virus, Mycobacterium leprae, Mycobacterium tuberculosis / Mycobacterium africanum, Mycobacterium bovis, Neisseria meningitidis, norovirus, poliovirus, Pseudomonas aeruginosa, rabies virus, Rickettsia prowleri, rotavirus, rubella virus Salmonella paratyphi, Salmonella typhi, Schistosoma, Shigella, Taenia solium, Taenia solium, Trichuris, Trypanosoma buderi, Trypanosoma congoense, Trypanosoma nigra, Trichinella spiralis, Varicella-zoster virus, Vibrio cholerae O1 and O139, Yersinia enterocolitica, Yersinia pestis, Treponema pallidum, HIV, Echinococcus, Plasmodium, Toxoplasma gondii, Streptococcus pneumoniae and / or Staphylococcus aureus.

[0114] Brucella spp. can include Brucella abortus, Brucella martatus, Brucella suis, and / or Brucella canis. Campylobacter spp. can include Campylobacter jejuni and / or Campylobacter coli. Legionella spp. can include Legionella pneumophila, Legionella medrelli, and / or Legionella longboast. Shigella spp. can include Shigella dysenteriae, Shigella flexneri, and / or Shigella boydii. Echinococcus spp. can include Echinococcus granulosus, Echinococcus multilocularis, Echinococcus voyensis, and / or Echinococcus septum. Plasmodium spp. can include Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale, Plasmodium malariae, and / or Plasmodium norocioli.

[0115] This yields the following advantages: the detection method can accurately and specifically detect a variety of pathogens, and therefore can be flexibly used to diagnose a variety of diseases. Attached Figure Description

[0116] Other advantages, features, and details of the invention are derived from the following detailed description of various embodiments of the invention with reference to the accompanying drawings. Here, the features mentioned in the claims and specification may be important to the invention individually or in any combination. The invention will be explained in detail with reference to the accompanying drawings, which are schematically illustrated herein: Figure 1 A view of a sample container used for detecting pathogens and / or cells within a sample is shown. Figure 2 A view showing a sandwich structure including the pathogen and / or cells to be detected within the sample. Figure 3 This diagram illustrates a detection system for detecting pathogens and / or cells within a sample. Figure 4 This shows a view of pathogens and / or cells enriched in the enrichment area of ​​the sample container. Figure 5 A schematic diagram of a detection method for detecting pathogens and / or cells within a sample is shown. Detailed Implementation

[0117] In the following figures, the same technical features are referred to by the same reference numerals even in different embodiments.

[0118] Figure 1 A sample container 10 with a sample processing unit 12 is shown, wherein the sample processing unit 12 includes a receiving area 13, a preparation area 14, and / or a mixing area 15. The receiving area 13 may refer to the region where the sample is introduced into the sample container 10. The preparation area 14 may refer to the region where the sample is pre-treated. The mixing area 15 may refer to the region where the sample is preferably mixed by motion. For this purpose, the mixing area 15 may preferably have a mixing structure 16. The mixing structure 16 may preferably include mixing elements 16a, in particular beads. Alternatively or additionally, the mixing structure 16 can be designed in the form of mixing elements 16b, in particular protrusions, as flaps and / or edges and / or mound-like protrusions arranged on the inner wall of the sample container 10 within the mixing area 15.

[0119] In addition, the sample container 10 includes an enrichment zone 17. The enrichment zone 17 may refer to the area where pathogens and / or cells 11 contained in the sample are concentrated.

[0120] The receiving region 13, the preparation region 14, the mixing region 15, and the enrichment region 17 may also form a single region, wherein these regions 13, 14, 15, and 17 preferably completely overlap.

[0121] Sample container 10 may refer to a reaction vessel. Sample container 10 is preferably made at least partially of polypropylene and / or a cyclic olefin copolymer (ethylene-norbornene copolymer). Additionally, sample container 10 preferably has a conical shape. Sample container 10 particularly includes a plurality of walls, wherein the angles of these walls preferably gradually narrow downwards.

[0122] The bottom 24 of the sample container 10 is preferably planar. Furthermore, the bottom 24 of the sample container 10 is particularly constructed with optically transparent properties. This allows for easy detection of pathogens and / or cells 11 contained in the sample through the bottom 24 of the sample container 10.

[0123] Figure 2 The sandwich structure 18 is shown, which is specifically constituted by an enriching agent 19, a staining agent 20, and pathogens and / or cells 11. The enriching agent 19 preferably comprises an antibody 21a. The staining agent 20 preferably comprises another antibody 21b. The antibody 21a preferably comprises a magnetic trapping agent 22, wherein the magnetic trapping agent 22 may be specifically designed as magnetic microparticles. The other antibody 21b preferably comprises a labeling agent 23, wherein the labeling agent 23 may be designed as a fluorescent labeling agent. The antibody 21a can bind to one target structure on the surface of the pathogen and / or cells 11, while the other antibody 21b can bind to a preferred other target structure on the surface of the pathogen and / or cells 11. Thus, the sandwich structure 18 can be generated by the enriching agent 19, the staining agent 20, and the pathogen and / or cells 11.

[0124] Figure 3 A detection system 100 for detecting pathogens and / or cells 11 within a sample is shown. The detection system 100 includes a sample container 10 containing a sample containing pathogens and / or cells 11. Furthermore, the detection system 100 includes a mixing unit 101 having 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 reagents pre-presented in the sample container 10 with the sample in the sample container 10. The reagents preferably include 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 staining agent 20 and / or at least one stabilizer. The mixing unit 101 may preferably include an eccentric mixer. Furthermore, the mixing unit 101 may particularly have a rotary mixer and / or a vibratory mixer.

[0125] Additionally, the detection system 100 includes an evaluation unit 103 designed to enrich pathogens and / or cells 11 within an enrichment zone 17 of the sample container 10, and to visualize and detect at least one pathogen 11 and / or at least one cell 11 enriched within the enrichment zone 17. The evaluation unit 103 includes a magnet 105 preferably used for enriching 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, bound to an enriching agent 19 and a staining agent 20. The enriching agent 19 preferably includes at least one antibody 21a with at least one magnetic trapping agent 22, particularly magnetic microparticles. The staining agent 20 preferably includes another antibody 21b with a labeling agent 23, particularly a fluorescent labeling agent. Magnet 105 is preferably positioned below the second sample holder 104 to ensure that pathogens and / or cells 11 contained in the sample are enriched in the enrichment area 17 of the sample container 10, wherein the enrichment area 17 is positioned close to the bottom 24 of the sample container 10, particularly directly at the bottom 24.

[0126] Additionally, the evaluation unit 103 may include an image processing unit 106, which preferably includes a fluorescence microscope 107. With the aid of the image processing unit 106, pathogens and / or cells 11 within the sample can be visualized and detected individually, preferably as fluorescent units.

[0127] Additionally, the evaluation unit 103 may include a calculation unit 108. The calculation unit 108 may be designed to quantify at least one pathogen 11 visualized by the image processing unit 106 and / or at least one cell 11 visualized by the image processing unit 106. Alternatively or additionally, the calculation unit may be designed to perform pattern recognition, wherein the Z-axis overlay sequence acquired by the image processing unit 106 is processed into a single image using maximum intensity projection, and pattern recognition is performed on the visualized and detected pathogens and / or cells based on this image. In particular, size-based filtering may be performed in the pattern recognition, wherein objects visible in the image that are all outside a preset size range, preferably larger than or smaller than 4 pixels, and / or whose shape exceeds a preset range, preferably a 2×2 pixel shape, are preferably filtered out.

[0128] In addition, the evaluation unit 103 may have a communication interface 109, which is preferably designed to transmit analysis data to a mobile device 110, especially a smartphone and / or a database 111, especially a data cloud, in a wired and / or wireless manner, preferably via Bluetooth and / or WLAN.

[0129] The evaluation unit 103 may also have a display device 112, especially a display, for displaying analysis data to the user.

[0130] Figure 4 A plan view of the bottom 24 of the sample container 10 is shown, in which individual pathogens and / or individual cells 11 within the enrichment area 17 of the sample container 10 are visible, particularly in the form of fluorescent units, and individual detection of pathogens and / or cells 11 is possible. Based on individual detection, quantification and / or pattern recognition of pathogens and / or cells 11 can be performed, in particular. The arrangement of pathogens and / or cells 11 is preferably similar to a starry sky.

[0131] Figure 5 A detection method 200 for detecting pathogens and / or cells 11 in a sample is shown. The detection method 200 includes, in a first step, introducing a sample, particularly a blood sample, into a receiving area 13 of a sample processing unit 12 of a sample container 10, wherein the sample contains pathogens and / or cells 11. Another step of the detection method 200 includes preparing the sample 202 in a preparation area 14 of the sample processing unit 12 of the sample container 10. In another step, the detection method 200 includes mixing the sample 203 in a mixing area 15 of the sample processing unit 12 of the sample container 10 by placing the sample container 10 into a mixing unit 101. A further step of the detection method 200 includes enriching the pathogens and / or cells 11 in the sample 204 in an enrichment area 17 of the sample container 10 by placing the sample container 10 into an evaluation unit 103. The final step of the detection method 200 includes evaluating the enrichment region 17 of the sample container 10 by means of the evaluation unit 103, wherein pathogens and / or cells 11 within the sample are visualized and detected by means of the evaluation unit 103.

[0132] Preferably, the sample container 10 contains pre-stored reagents, particularly lyophilized and / or dried reagents. The reagents preferably include at least one lysing agent and / or at least one permeabilizing agent and / or at least one enriching agent 19 and / or at least one staining agent 20 and / or at least one stabilizer. A sample, particularly a blood sample, can then be added to the reagents. The enriching agent 19 may preferably have an antibody 21a linked to a magnetic trapping agent 22, particularly magnetic microparticles. The staining agent 20 may preferably include an antibody 21b having a labeling agent 23, particularly a fluorescent labeling agent. The stabilizer may particularly have a polysaccharide, preferably dextran, to maintain the functionality of the sample and / or reagents.

[0133] Sample container 10, containing, in particular, lyophilized and / or dried reagents and samples, can preferably be fed into a first sample holder 102 of a mixing unit 101, particularly having an eccentric mixer. Mixing of the sample and / or reagents can preferably be performed via the mixing unit 101, wherein, during mixing, in particular, lysis and / or permeation of pathogens and / or cells 11 within the sample, and / or enrichment of pathogens and / or cells 11 within the sample container 10 in the enrichment zone 17, and / or labeling of pathogens and / or cells 11 within the sample can be performed.

[0134] It is also conceivable that the sample container 10 is subsequently fed into a second sample holder 104 of the evaluation unit 103. The evaluation unit 103 may preferably include a magnet 105 and / or a fluorescence microscope 107. Pathogens and / or cells 11 are preferably bound to an enriching agent 19 having an antibody 21a, wherein the antibody 21a is particularly attached to a magnetic trapping agent 22. By means of the magnet 105, pathogens and / or cells 11 can thus be enriched within the enrichment zone 17 by magnetic force. The enriched pathogens and / or cells 11 are also bound, in particular, to a staining agent 20 having an antibody 21b, wherein the antibody 21b is particularly attached to a labeling agent 23, especially a fluorescent labeling agent. Thus, pathogens and / or cells 11 can be visualized and detected by means of the fluorescence microscope 107.

[0135] This enables highly sensitive and precise detection of cells and / or pathogens 11 in samples, especially blood samples, including individual pathogens and / or individual cells 11. Therefore, it can also detect extremely low concentrations of pathogens and / or individual cells 11 within a sample and determine the precise increase or decrease in the number of pathogens and / or individual cells 11 within the sample.

[0136] The above description of the accompanying drawings illustrates the invention within the scope of the examples. It is obvious that the various features of these embodiments can be freely combined with each other as long as they are technically reasonable, without departing from the scope of the invention.

[0137] List of reference numerals 10 sample containers 11 Pathogens and / or cells 12 sample processing units 13 Reception Area 14 Preparation Zone 15 Mixed Zones 16 Hybrid Structure 17 rich areas 18-sandwich structure 19 enriching agents 20 staining agents 21a antibody 21b antibody 22 Magnetic trapping agent 23 markers 24 bottom 100 Detection System 101 Hybrid Unit 102 First Sample Stent 103 Evaluation Unit 104 Second Sample Stent 105 magnet 106 Image Processing Units 107 microscope 108 computing units 109 communication interface 110 mobile devices 111 Database 112 display device 200 detection method 201 Introduction 202 Preparation 203 Mixed 204 enrichment 205 assessment

Claims

1. A sample container (10) for detecting pathogens and / or cells (11) in a sample, particularly a blood sample, the sample container (10) having a sample processing unit (12) comprising: - At least one receiving area (13) of the sample processing unit (12) for receiving samples. - At least one preparation area (14) of the sample processing unit (12) for preparing a sample. - At least one mixing region (15) of the sample processing unit (12) for mixing samples, wherein the mixing region (15) includes a mixing structure (16), and - At least one enrichment zone (17) for enriching pathogens and / or cells (11) contained in the sample.

2. The sample container (10) according to claim 1, wherein, The enrichment region (17) includes at least one enriching agent (19) and at least one staining agent (20), wherein the enriching agent (19) includes at least one antibody (21a) having at least one magnetic trapping agent (22), particularly magnetic microparticles, for capturing pathogens and / or cells (11) in the sample, wherein the at least one staining agent (20) includes at least one other antibody (21b) having at least one labeling agent (23), particularly fluorescent labeling agent, for labeling pathogens and / or cells (11) in the sample, wherein preferably a sandwich structure (18) can be generated by the at least one enriching agent (19) and the at least one staining agent (20).

3. The sample container (10) according to any one of the preceding claims, wherein, The sample volume includes between 1 µL and 1 mL, preferably between 10 and 100 µL, and particularly preferably between 20 and 30 µL.

4. The sample container (10) according to any one of the preceding claims, wherein, The enrichment area (17) is arranged near the bottom inside the sample container (10), wherein, in particular, at least one bottom (24) of the sample container (10) is constructed with optical-grade transparency.

5. The sample container (10) according to any one of the preceding claims, wherein, The preparation area (14) has at least one lysing agent and / or at least one permeabilizing agent for lysing and / or permeating the pathogen and / or cells (11).

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

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

8. The detection system (100) according to claim 6 or 7, wherein, The evaluation unit (103) includes at least one magnet (105), wherein the evaluation unit (103) is designed to enrich at least one pathogen and / or at least one cell (11) in an enrichment zone (17) of the sample container (10) by means of the magnet (105), wherein the at least one pathogen and / or the at least one cell (11) is bound to the at least one enriching agent (19), wherein the enriching agent (19) includes at least one antibody (21a) having a magnetic trapping agent (22), in particular magnetic particles, wherein the enrichment zone (17) is arranged in the sample container (10) particularly near the bottom.

9. The detection system (100) according to any one of claims 6 to 8, wherein, The evaluation unit (103) is designed to visualize and detect the pathogens and / or cells (11) within the enrichment area (17) of the sample container (10) through the bottom (24) of the sample container (10), wherein the bottom (24) is preferably constructed with optical-grade transparency, and / or the evaluation unit (103) is designed to detect individual pathogens and / or individual cells (11) within the enrichment area (17) of the sample container (10).

10. The detection system (100) according to any one of claims 6 to 9, wherein, The evaluation unit (103) includes an image processing unit (106) designed to visualize the pathogen and / or cells (11) individually, wherein the image processing unit (106) includes, particularly preferably, an inverted and / or automated fluorescence microscope (107) designed to visualize the pathogen and / or cells (11) as fluorescent units.

11. The detection system (100) according to any one of claims 6 to 10, wherein, The evaluation unit (103) includes a computing unit (108) which is designed to quantify and / or perform pattern recognition on at least one visualized pathogen and / or at least one visualized cell (11).

12. The detection system (100) according to any one of claims 6 to 11, wherein, The evaluation unit (103) includes a communication interface (109) designed to transmit the analysis data of the evaluation unit (103) to a mobile device (110) and / or a database (111), particularly a data cloud, and / or the evaluation unit (103) includes a display device (112), particularly a display, designed to display the analysis data of the evaluation unit (103) to a user.

13. A detection method (200) for detecting pathogens and / or cells (11) in a sample, particularly a blood sample, preferably using a detection system (100) according to any one of claims 6 to 12, comprising the following steps: - The receiving area (13) of the sample processing unit (12) of the sample container (10) is used to introduce a sample, especially a blood sample, into the sample container (201), wherein the sample contains pathogens and / or cells (11). - The sample is prepared (202) in the preparation area (14) of the sample processing unit (12) of the sample container (10). - By feeding the sample container (10) into the mixing unit (101), the sample is mixed (203) in the mixing area (15) of the sample processing unit (12) of the sample container (10). - By sending the sample container (10) into the evaluation unit, the pathogens and / or cells (11) in the sample are enriched (204) in the enrichment area (17) of the sample container (10). - The enrichment area (17) of the sample container (10) is evaluated (205) by the evaluation unit (103), wherein pathogens and / or cells (11) within the sample are visualized and detected by means of the evaluation unit.

14. The detection method (200) according to claim 13, wherein, The preparation of the sample (202) includes the lysis and / or permeation of the pathogen and / or cells (11), and / or the enrichment (204) of the pathogen and / or cells (11) in the enrichment zone (17) of the sample container includes magnetic capture and / or fluorescent labeling, and / or The evaluation (205) of the enriched region includes the quantification or pattern recognition of the pathogen and / or cells (11), wherein the enriched region (17) is disposed particularly close to the bottom within the sample container (10), and the evaluation (205) of the enriched region (17) is performed particularly through the bottom (24) of the sample container (10), wherein the bottom (24) is constructed with optically grade transparency.

15. The detection method (200) according to claim 13 or 14, wherein, The pathogen (11) includes at least one malaria pathogen, particularly Plasmodium falciparum and / or Plasmodium vivax and / or Plasmodium ovale and / or Plasmodium norotri and / or Plasmodium malariae, and / or the cell (11) includes at least one red blood cell.

16. The detection method (200) according to claim 13 or 14, wherein, The pathogen (11) includes at least one human immunodeficiency virus, and / or the cell (11) includes at least one lymphocyte, preferably a helper T cell, particularly preferably a CD4-T cell and / or a CD8-T cell.

17. The detection method (200) according to any one of claims 13 to 16, used for diagnosing at least one disease, wherein, The diseases mentioned are especially those caused by pathogens (11), particularly fungi, viruses and / or bacteria and / or pathogens, especially at least one of the following groups: adenovirus, hookworm, ascarid, babesi, Bacillus anthracis, Bordetella pertussis, Bordetella parapertussis, Treponema relapsingis, Brucella, Campylobacter, tapeworms, Chlamydia psittaci, Clostridium botulinum, Corynebacterium diphtheriae, Coxsella beniformis, Cryptosporidium spp., Ebola virus, Echinococcus multilocularis, Echinococcus granulosus, Escherichia coli, enterohemorrhagic Escherichia coli (EHEC), E. eukaryotes, Tulafrancsis, tick-borne encephalitis virus, yellow fever virus, Giardia lamblia, Haemophilus influenzae, Hantavirus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus, hepatitis E virus Viruses, influenza virus, Lassa virus, Legionella spp., human pathogenic Leptospira spp., Listeria monocytogenes, Marburg virus, measles virus, mumps virus, Mycobacterium leprae, Mycobacterium tuberculosis / Mycobacterium africanum, Mycobacterium bovis, Neisseria meningitidis, norovirus, poliovirus, Pseudomonas aeruginosa, rabies virus, Rickettsia prowleri, rotavirus, rubella virus, paracetamol. Salmonella typhi, Salmonella typhi, Schistosoma, Shigella, Taenia solium, Taenia solium, Trichuris, Trypanosoma, Trypanosoma congoense, Trypanosoma nigra, Trichinella spiralis, Varicella-zoster virus, Vibrio cholerae O1 and O139, Yersinia enterocolitica, Yersinia pestis, Treponema pallidum, HIV, Echinococcus, Plasmodium, Toxoplasma gondii, Streptococcus pneumoniae and / or Staphylococcus aureus.

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