Combined test card and combined test method for immunodetection and cell analysis

Abstract

The application provides a combined detection card for immune detection and cell analysis and a combined detection method. The combined detection card integrates an immune detection component and a cell analysis component together through a shell structure formed by a detection card bottom shell matched with a detection card upper cover. The shell is provided with a first sample adding hole and a second sample adding hole for respectively adding samples to the immune detection component and the cell analysis component, a first reading window for exposing an immune detection area of the immune detection component, and a second reading window and a third reading window for exposing a detection surface of the cell analysis component. The combined detection card is matched with a combined detection device to simultaneously realize immune detection and cell analysis functions, thereby providing a basic condition for POCT level synchronous immune and cell on-site combined detection. The cell analysis component of the combined detection card is provided with a sample adding gap, a vent hole, one or multiple test areas with different thicknesses, so that bubble generation can be avoided, accurate control of a cell sample amount can be realized, and the precision of cell counting and typing detection can be simultaneously considered.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical detection technology, and in particular relates to a combined detection card for immune detection and cell analysis and a combined detection method. Background Art

[0002] In vitro diagnostics (IVDs) are at the core of evidence-based Western medicine. They analyze patient fluids such as blood, urine, and saliva to determine a patient's current physical condition, providing objective, clear, and quantitative evidence for physicians' diagnoses. With the advancement of technology, IVDs have gradually expanded from clinical laboratories and central laboratories to emergency departments, ICUs, and other clinical departments in large hospitals, as well as mobile medical devices like ambulances and medical clinics, and even into homes. This has led to the concept of point-of-care testing (POCT), which allows non-professionals to quickly obtain test results on-site, using non-specialized laboratories. POCT-related technology products, with their professional performance and flexible operation, provide crucial support for the establishment of a national tiered diagnosis and treatment system, while also enabling personalized, immediate health management and preventive treatment.

[0003] For body fluid samples such as blood and urine, the detection targets mainly include specific disease-related proteins in the sample matrix and human cells. Generally, for the detection of specific proteins, the mainstream technology is immunoassay based on immunochromatographic technology. That is, based on the specific recognition of antigens and antibodies, the protein adsorption capacity of solid-phase materials such as nitrocellulose membranes is used as a carrier for the fixation of antigen-antibody probes. The capillary siphon force of solid-phase materials such as nitrocellulose membranes is used as the driving force for the surge and reaction of liquid samples. The specific recognition of antigens and antibodies is quantitatively demonstrated through the optical, electrical, and magnetic signals of biomarkers coupled to antigens and antibodies. For human cell analysis, commonly used instruments are flow cytometers and fully automatic hematology analyzers. In terms of function, both are mainly based on cell count measurement (i.e., "counting") and type differentiation (i.e., "typing").

[0004] Among them, the core principle of the flow cytometer is to use the sample tube and sheath fluid tube of the liquid flow system to make the sheath fluid surround the sample to be tested and flow at high speed in a circular stream, so as to arrange the cells to be tested in a single row and pass through the laser irradiation area in sequence. By analyzing the forward scattered light, side scattered light, and side fluorescence of each cell one by one, the number and type information of the cells can be finally obtained.

[0005] The primary counting principle used in fully automated hematology analyzers is the Coulter principle (also known as the electrical impedance method). This principle states that when cells suspended in an electrolyte flow through a small orifice tube, they displace an equal volume of electrolyte. This causes a transient change in the resistance between the inner and outer electrodes of the tube in a constant current circuit, generating potential pulses whose magnitude and frequency are proportional to the size and number of cells, thereby achieving cell counting. Prior to the Coulter method, selective hemolytic agents or chemical reagents are used to disrupt other cells while preserving the integrity of specific cells. This, combined with the counting function of the Coulter method, allows for both cell typing and counting.

[0006] Both methods analyze single cells and can therefore achieve accurate cell counting and typing. However, both methods suffer from the problems of expensive instruments, large size, and specialized operation, making them unsuitable for situations where cell analysis is required on-site or in non-specialized settings.

[0007] To solve the above problems, in clinical testing applications, existing cell analysis detection equipment uses disposable counting slides for sampling. The core structure of the counting slide is an open measurement cavity formed by two parallel transparent thin slices at a certain distance and a bottom. The measurement cavity has a measurement area with a thickness further reduced to 100 μm. The measurement cavity has the following functions: (1) Reagent fixation: red blood cell hemolysis agents (quaternary ammonium salts, saponins, etc.) and white blood cell color dyes (hematoxylin, methylene blue, etc.) are fixed on the inner wall of the measurement cavity; (2) Blood processing: The blood sample enters the measurement cavity by active aspiration, and red blood cell hemolysis and white blood cell staining are completed in the measurement cavity; (3) Cell spreading: The thickness of the detection area is further reduced, so that the white blood cells in the sample are spread as much as possible in a single layer in the measurement area, which is convenient for single cell analysis; (4) Detection connection: The detection system can obtain images of the single-layer spread cells in the measurement area through the side wall of the measurement cavity. Therefore, cell counting and typing based on cell morphology recognition are achieved by relying on color dyes.

[0008] Currently, immunoassays and cell analysis (including cell counting and typing analysis) are generally performed separately during clinical testing. Using two different instruments for testing results in low efficiency, high cost, and a large amount of blood sampling. Improving testing efficiency and speed is crucial, especially in emergency situations, to reducing patient waiting time and physician workload.

[0009] At present, there is no POCT-level combined detection instrument for immunoassay and cell analysis. In view of the urgent demand for POCT-level combined detection instrument for immunoassay and cell analysis, it is necessary to design and develop detection cards for immunoassay and cell analysis to meet the needs of on-site combined detection of immunoassay and cell. Summary of the Invention

[0010] To solve the above problems, the present invention provides a combined detection card for immunoassay and cell analysis.

[0011] The technical solution adopted in the present invention is:

[0012] A combined test card for immunoassay and cell analysis comprises: a test card upper cover (100), a test card bottom shell (200), and an immunoassay component (300) and a cell analysis component (400) installed in a housing formed by the test card upper cover (100) and the test card bottom shell (200), wherein:

[0013] The detection card bottom shell (200) is provided with a first placement groove (21) for installing the immune detection component (300), a second placement groove (22) for installing the cell analysis component (400), and a third interpretation window (23) for viewing the detection surface of the cell analysis component (400);

[0014] The detection card upper cover (100) is provided with a first sample loading hole (16) for loading a sample into the immune detection component (300), a second sample loading hole (17) for loading a sample into the cell analysis component (400), a first interpretation window (18) for viewing the immune detection area of ​​the immune detection component (300), and a second interpretation window (19) for viewing the detection surface of the cell analysis component (400).

[0015] The first interpretation window (18) is a window hole that passes through the upper cover (100) of the detection card, and the second interpretation window (19) and the third interpretation window (23) are window holes that pass through the upper cover (100) of the detection card and the bottom shell (200) of the detection card, respectively, and the second interpretation window (19) and the third interpretation window (23) are aligned.

[0016] In the above-mentioned combined detection card for immunoassay and cell analysis, the first sample loading hole (16) and the second sample loading hole (17) are connected.

[0017] In the above-mentioned combined test card for immunoassay and cell analysis, the cell analysis component (400) comprises an assembly handle (1) and a test cavity (2) provided on the assembly handle (1), wherein the test cavity (2) is a semi-open cavity formed by two parallel cavity side walls (3) with a certain gap, and comprises a test area (4), an injection port (5), and a drainage area (6) connecting the injection port (5) and the test area (4), wherein the thickness of the test area (4) is Less than the thickness of the drainage area (6) The injection port (5) is aligned with the second sample addition hole (17), and the test area (4) is aligned with the second interpretation window (19) and the third interpretation window (23).

[0018] In the above-mentioned combined test card for immunoassay and cell analysis, at least one exhaust hole (7) is provided on the test cavity (2), and the exhaust hole (7) is a through hole connecting the interior of the test cavity (2) with the outside atmosphere, and the exhaust hole (7) penetrates one cavity side wall of the test area (4) or the drainage area (6) or symmetrically penetrates two cavity side walls of the test area (4) or the drainage area (6); the exhaust hole (7) is preferably an inverted cone-shaped through hole, with the small end of the cone opening facing the inside of the test area (4) or the drainage area (6) and the large end of the cone opening facing the outside of the test cavity (2); the exhaust hole (7) is preferably connected to the outside atmosphere through a connecting hole (20) provided on the test card upper cover (100) and / or the test card bottom shell (200).

[0019] In the above-mentioned combined test card for immunoassay and cell analysis, one or more test areas (4) are provided in the test cavity (2), and the thickness of one test area (4) is The range is 60μm-120μm; multiple test areas (4) are independent and connected to each other, with the same or different thicknesses, and the thickness of any test area (4) is less than the thickness of the drainage area (6) , the thickness of any test area (4) ranges from 60μm to 120μm, and the thickness of the drainage area (6) The range is 120μm-500μm.

[0020] In the combined test card for immunoassay and cell analysis, the test cavity (2) is provided with two test areas (4) of different thicknesses, namely a first test area (41) and a second test area (42), and the first test area (41) and the second test area (42) are connected by a drainage area (6).

[0021] In the above-mentioned combined test card for immunoassay and cell analysis, the sample injection port (5) is located at the upper edge opening of the two cavity side walls (3) of the test cavity (2), wherein the upper edge of one of the two cavity side walls (3) located at the sample injection port (5) is provided with a sample injection notch (8) so as to inject the liquid sample through the sample injection notch (8); the sample injection notch (8) is opposite to the second sample addition hole (17).

[0022] In the above-mentioned combined detection card for immunoassay and cell analysis, the injection port (5) is in a concave arc shape, and the angle α between the tangent of the downward arc and the horizontal reference plane of the injection port (5) is in the range of 15°-45°.

[0023] In the combined detection card for immunoassay and cell analysis, the inner edge of the end edge of the cavity side wall (3) is provided with a transition fillet (10), and the transition fillet value R ranges from 0.2 mm to 1.5 mm.

[0024] In the above-mentioned combined detection card for immunoassay and cell analysis, the immunoassay component (300) is an immunochromatographic test paper, comprising a sample pad (11), a conjugation pad (12), an analysis membrane (13), an absorbent paper (14) and an adhesive backing (15). The sample pad (11), the conjugation pad (12), the analysis membrane (13) and the absorbent paper (14) are sequentially adhered and fixed to the adhesive backing (15) in an overlapping manner. The sample pad (11) is aligned with the first sample addition hole (16), and the analysis membrane (13) is aligned with the first reading window (18).

[0025] The present invention also provides a combined detection method using any of the above-mentioned combined detection cards for immunoassay and cell analysis, comprising the following steps:

[0026] S1) adding liquid sample to the sample well of the combination test card;

[0027] S2) waiting for a predetermined time until an immune detection area is formed on the analysis membrane (13) of the immune detection component (300) and a detection surface is formed on the test area (4) of the cell analysis component (400);

[0028] S3) Using an immunoassay instrument and a cell analysis instrument, respectively, perform immunoassay on the immunoassay area of ​​the immunoassay component of the combination test card and perform cell analysis on the detection surface of the cell analysis component.

[0029] In the above-mentioned combined detection method, the sample addition operation in step S1) is as follows:

[0030] Aspirate the liquid sample with a pipette, place the pipette on the first sample loading hole (16) and the second sample loading hole (17) of the combined detection card, and add the sample so that the liquid sample flows into the sample pad (11) of the immunoassay component (300) and the sample inlet (5) of the cell analysis component (400), respectively; or

[0031] The liquid sample is added to the connected first sample addition well (16) or the second sample addition well (17) at one time through the pipette, so that the liquid sample flows into the sample pad (11) of the immunoassay component (300) and the sample inlet (5) of the cell analysis component (400) respectively.

[0032] The beneficial effects of the present invention are as follows: the combined detection card of the present invention integrates the immune detection component and the cell analysis component through the shell structure formed by the detection card bottom shell and the detection card upper cover, and can be used in a combined detection device provided with an immune detection unit and a cell detection and analysis unit. While completing the realization of immune detection and cell analysis, it provides the basic conditions for realizing synchronous immune and cell on-site joint detection at the POCT level; the cell analysis component of the combined detection card can avoid the generation of bubbles by providing a sampling notch, an exhaust hole, and one or multiple test areas of different thicknesses, thereby achieving precise control of the cell sample amount and taking into account the precision of cell counting and typing detection at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1A 1 is a schematic plan view of the structure of the cell analysis component in the combined detection card of the present invention;

[0034] Figure 1B This is a schematic diagram of the three-dimensional structure of the cell analysis component in the combined detection card of the present invention;

[0035] Figure 1C yes Figure 1A The cross-sectional view taken along line AA;

[0036] Figure 2A 1 is a schematic plan view of the structure of the cell analysis component in the combined detection card of the present invention;

[0037] Figure 2B This is a schematic diagram of the three-dimensional structure of the second embodiment of the cell analysis component in the combined detection card of the present invention;

[0038] Figure 2C yes Figure 2A A cross-sectional view taken along line BB;

[0039] Figure 3A 1 is a schematic plan view of the structure of the cell analysis component in the combined detection card of the present invention;

[0040] Figure 3B This is a schematic diagram of the three-dimensional structure of the cell analysis component in the combined detection card of the present invention;

[0041] Figure 3C yes Figure 3A A cross-sectional view taken along line C1-C1;

[0042] Figure 3D yes Figure 3A A cross-sectional view taken along line C2-C2;

[0043] Figure 4A 1 is a schematic diagram of the three-dimensional structure of the first embodiment of the combined detection card of the present invention;

[0044] Figure 4B 1 is a perspective schematic diagram of a first embodiment of the combined detection card of the present invention;

[0045] Figure 4C 1 is a front structural diagram of a first embodiment of the combined detection card of the present invention;

[0046] Figure 4D This is a schematic diagram of the reverse structure of the first embodiment of the invented combined detection card;

[0047] Figure 5A This is a schematic diagram of the three-dimensional structure of the second embodiment of the combined detection card of the present invention;

[0048] Figure 5B This is a perspective schematic diagram of a second embodiment of the combined detection card of the present invention;

[0049] Figure 5C This is a schematic diagram of the front structure of the second embodiment of the combined detection card of the present invention;

[0050] Figure 5D This is a schematic diagram of the reverse structure of the second embodiment of the invented combined detection card;

[0051] Figure 6 This is an example of the structure of the immunoassay component in the combined test card of the present invention.

[0052] The reference numerals in the figures are as follows:

[0053] 100-test card cover, 200-test card bottom shell, 300-immunoassay component, 400-cell analysis component;

[0054] 1-Assembly handle, 2-Test cavity, 3-Cavity side wall;

[0055] 4-test area, 41-first test area, 42-second test area;

[0056] 5-inlet, 6-drainage area;

[0057] 7-exhaust hole, 71-first exhaust hole, 72-second exhaust hole, 73-third exhaust hole, 74-fourth exhaust hole;

[0058] 8-injection notch, 9-transition fillet, 10-liquid bridge surface;

[0059] 11-sample pad, 12-conjugate pad, 13-analytical membrane, 14-absorbent paper, 15-adhesive backing;

[0060] 16-first sample loading well, 17-second sample loading well, 18-first reading window, 19-second reading window;

[0061] 20-connecting hole, 201-first connecting hole, 202-second connecting hole, 203-third connecting hole, 204-fourth connecting hole;

[0062] 21 - first placement slot, 22 - second placement slot, 23 - third interpretation window. DETAILED DESCRIPTION

[0063] In order to solve the problem that existing in vitro diagnostics lack POCT-level combined immune and cell testing technology, and to avoid the problems of cell analysis being unable to simultaneously take into account the precision of cell counting and typing, being prone to bubble interference, and having low sampling accuracy, the present invention provides a combined test card for immune detection and cell analysis and a combined detection method. The combined test card includes a test card cover, a test card bottom shell, and an immune detection component and a cell analysis component installed in a shell formed by the test card cover and the test card bottom shell.

[0064] The cell analysis component includes an assembly handle and a test cavity provided on the assembly handle. The test cavity is a semi-open cavity formed by two parallel cavity side walls with a certain gap. The thickness of the test area is determined by the design of an injection port with an injection notch, one or multiple test areas of different thicknesses, a drainage area for connecting the injection port and the test area, and an exhaust hole for removing bubbles. Smaller than the thickness of the drainage area This structure enables rapid and smooth injection of liquid samples, avoids the generation of bubbles, and achieves precise control of cell sample volume while simultaneously ensuring the precision of cell counting and typing detection, laying the foundation for accurate on-site cell analysis. The immunoassay component is an immunochromatographic test strip, including a sample pad, conjugate pad, analytical membrane, absorbent paper, and an adhesive backing.

[0065] The test card base is equipped with a first placement slot for mounting the immunoassay component, a second placement slot for mounting the cell analysis component, and a third interpretation window facing the test area of ​​the cell analysis component. The test card cover is equipped with a first sampling hole for loading samples into the immunoassay component, a second sampling hole for loading samples into the cell analysis component, a first interpretation window facing the analysis membrane of the immunoassay component, and a second interpretation window facing the test area of ​​the cell analysis component. This combined test card integrates the immunoassay component and the cell analysis component through the shell structure formed by the test card base and the test card cover. When used with a combined detection device equipped with an immunoassay unit and a cell detection and analysis unit, it can simultaneously perform immunoassay and cell analysis functions, providing the basic conditions for achieving synchronous on-site immunoassay and cell analysis at the point-of-care (POCT) level.

[0066] The combined detection card for immunoassay and cell analysis and the combined detection method of the present invention are described in detail below with reference to the examples and the accompanying drawings.

[0067] Example 1

[0068] Cell Analysis Kit 400:

[0069] Figure 1A-1C This is an example of the structure of the cell analysis component of the present invention. Figure 1A-1C In the illustrated embodiment of the cell analysis assembly, the cell analysis assembly 400 includes an assembly handle 1 and a test cavity 2 disposed on the assembly handle 1. The rear end of the assembly handle 1 is a handheld portion designed to be handheld. For example, in this embodiment, the handheld portion is rectangular, and a test cavity 2 with an arc-shaped edge extends from the front end of the assembly handle 1. The test cavity 2 can be integrally formed with the assembly handle 1, or the test cavity 2 can be bonded to the front end of the assembly handle 1. The test cavity 2 is a semi-open cavity formed by two parallel cavity sidewalls 3 with a certain gap, and includes an injection port 5, an injection notch 8, a test area 4, and a drainage area 6 connecting the injection port 5 and the test area 4, wherein:

[0070] In this embodiment, the injection port 5 is located at the upper edge opening of the two cavity side walls 3 of the test cavity 2. The upper edge of one of the two cavity side walls 3 located at the injection port 5 is provided with an injection notch 8, which facilitates the positioning of the liquid sample pipette so that the sample can be injected through the injection notch 8. The injection port 5 is arc-shaped, and the angle α between the tangent line of its downward arc and the horizontal reference plane of the injection port 5 (see Figure 1A ), which determines the direction of flow of the liquid sample to be tested into the drainage area 6, thereby ensuring that the liquid sample to be tested spontaneously flows into the test area 4 in a predetermined manner and fills the test area 4. The preferred range of angle α is 15°-45°.

[0071] The test area 4 is located in the test cavity 2. The shape of the test area 4 can be a rectangle, square, trapezoid, circle or a combination of arcs and other shapes. Each shape can have rounded corners, right angles or a combination of rounded corners and right angles. The present invention does not limit the specific shape of the test area 4. The test area 4 has a single thickness. The liquid sample enters the test area 4 to form a detection surface. The test area 4 with a large thickness has a large sample carrying capacity per unit area of ​​the detection surface and a large depth of field, which is suitable for the overall accurate measurement of the number of cells; the test area 4 with a small thickness has a large spreading area of ​​the same volume of liquid sample on the detection surface, which is suitable for the precise distinction of cell types. The thickness of the test area 4 is 60μm-120μm.

[0072] The drainage area 6 is located in the test cavity 2 and is connected to the injection port 5 and the test area 4. The thickness of the drainage area 6 is The range is generally 120μm-500μm. Figure 2C As shown, the thickness of the test area 4 Less than the thickness of drainage area 6 The liquid sample enters the flow path formed by the drainage area 6 from the sampling port 5 and is evenly and quickly introduced into the test area 4 and fills it.

[0073] The thickness of the test zone 4 and the thickness of the drainage zone 6 determine the flow state and spreading state of the liquid sample in the drainage zone 6. The liquid sample enters the drainage zone 6 through the inlet 5. The sample is drawn into the drainage zone 6 during a pure inertial rise phase under the action of capillary force. The relationship between the volume of the drawn liquid sample and the thickness of the test zone 4 can be obtained using Equation 1) for the pure inertial rise phase of capillary flow:

[0074]

[0075] Thickness of test area 4 Less than the thickness of drainage area 6 , which is conducive to the discharge of bubbles. To ensure that the liquid can flow continuously from the drainage area 6 into the test area 4 under the action of capillary force and fill it, the capillary pressure is required to be greater than zero. Capillary force and the thickness of the test area 4 , the thickness of drainage area 6 , the surface tension of the liquid being tested, and the contact angle of the liquid being tested on the surface of the material in the drainage area 6 have the following relationship:

[0076]

[0077] From formula 2), we can know that by designing the thickness of the drainage area 6 and the thickness of test area 4 , it is possible to adjust different liquid flow rates and liquid laminar flow characteristics, thereby avoiding the formation of bubbles.

[0078] Specifically, in this embodiment, the test area 4 is provided with one, located in the test cavity 2, and having a single thickness. , its shape is a rounded rectangle. When the thickness of the test area 4 When the thickness of the test area 4 is larger, the depth of field of the detection surface formed on the side wall 3 of the cavity is large, and the sample carrying capacity per unit area is large, which is suitable for the overall accurate measurement of the number of cells. The thickness of the test area 4 is preferably 90μm-120μm; when the thickness of the test area 4 is When the thickness is smaller, the depth of field of the detection surface formed on the cavity side wall 3 is small and the sample spreading area per unit volume is large, which is suitable for precise differentiation of cell types. At this time, the thickness of the test area 4 is preferably 60 μm-90 μm.

[0079] The chamber sidewalls 3 can be used for subsequent cell counting and typing monitoring and analysis. To prevent the liquid sample from escaping due to gravity when the cell analysis assembly is moved, the inner edges of the chamber sidewalls 3 are provided with a transition radius 9, with a radius R ranging from 0.2mm to 1.5mm. When the test chamber 2 is filled with sample, the liquid sample forms a stable liquid bridge 10 at the transition radius 9 on the two chamber sidewalls 3, effectively balancing the liquid's gravity and preventing it from escaping.

[0080] The structural design of the first embodiment is suitable for applications where the number of liquid cells per unit volume is small or the type abundance is low, or where only liquid cell counting or liquid cell typing is required. According to the application, a test area 4 with a single thickness is selected and an appropriate thickness is set. , in order to perform high-precision measurement of cell number or high-precision analysis of single cell types.

[0081] Obviously, in this embodiment, a test area 4 may also have multiple thicknesses. 、 etc., and the thickness of each test area 4 is smaller than the thickness of the drainage area 6. The capillary force that drives the liquid sample to be tested into the test cavity 2 is proportional to the thickness of each test area 4 and the thickness of the drainage area. The relationship still satisfies Equation 2). The sample enters the test area 4 to form a detection surface with different depths of field and different spreading states, which can simultaneously take into account the precision measurement of cell counting and typing.

[0082] Immunoassay Kit 300:

[0083] like Figure 6 As shown, the immunoassay component 300 is an immunochromatographic test paper, comprising a sample pad 11, a conjugated pad 12, an analytical membrane 13, an absorbent paper 14 and an adhesive backing 15, wherein the sample pad 11, the conjugated pad 12, the analytical membrane 13, the absorbent paper 14 are sequentially adhered and fixed to the adhesive backing 15 in an overlapping edge manner, thereby ensuring the continuity of the flow of the liquid sample on the immunochromatographic test paper.

[0084] Combination Test Card:

[0085] See also Figures 4A to 4D , which is the structure of Example 1 of the combined immunoassay and cell analysis test card of the present invention, the combined test card includes a test card upper cover 100, a test card bottom shell 200, and an immunoassay component 300 and a cell analysis component 400 installed in a housing formed by the test card upper cover 100 and the test card bottom shell 200.

[0086] The test card bottom shell 200 is provided with a first placement slot 21 for installing the immune detection component 300, a second placement slot 22 for installing the cell analysis component 400, and a third interpretation window 23 facing the test area 4 of the cell analysis component; the test card cover 100 is provided with a first sample loading hole 16 for loading the immune detection component 300, a second sample loading hole 17 for loading the cell analysis component 400, a first interpretation window 18 facing the analysis membrane 13 of the immune detection component 300, and a second interpretation window 19 facing the test area 4 of the cell analysis component 400, wherein the first interpretation window 18 is a window hole passing through the test card cover 100, the second interpretation window 19 and the third interpretation window 23 are window holes passing through the test card cover 100 and the test card bottom shell 200 respectively, and the two window holes are aligned. The combined test card integrates the immune detection component 300 and the cell analysis component 400 through the shell structure formed by the test card bottom shell 200 and the test card upper cover 100. When used in conjunction with a joint detection device equipped with an immune detection unit and a cell detection and analysis unit, it can simultaneously realize immune detection and cell analysis functions, providing conditions for realizing synchronous on-site joint detection of immune and cells at the POCT level.

[0087] The assembly handle 1 of the cell analysis component 400 of this embodiment 1 is mounted in the second placement slot 22 on the test card base 200, and the immunoassay component 300 is placed in the first placement slot 21 on the test card base 200. After the test card cover 100 is closed with the test card base 200, the first sample loading hole 16 on the test card cover 100 faces the sample pad 11 of the immunoassay component 300, and the second sample loading hole 17 faces the sample inlet of the cell analysis component 400.

[0088] Combine Figure 6 During sample injection, the liquid sample is injected into the sample pad 11 of the corresponding immunoassay component 300 through the first sample injection hole 16, and enters the conjugate pad 12 through penetration and siphoning, so that the marker-bioactive molecule conjugate therein is redissolved and freed, and under the siphoning action of the absorbent paper 14, it leaves the conjugate pad 12 and enters the analysis membrane 13 to produce the immunoassay area, which is aligned with the first interpretation window 18 of the test card cover 100.

[0089] During sample injection, the liquid sample is injected into the corresponding sample inlet 5 of the cell analysis component 400 through the second sample injection hole 17. Under the action of capillary force, the liquid sample flows into the test area 4 through the drainage area 6. A detection surface is formed on the two cavity side walls 3 of the test area 4 of the cell analysis component 400. The detection surface is aligned with the second interpretation window 19 and the third interpretation window 23.

[0090] Specifically, in the above embodiment, the first sample loading hole 16 and the second sample loading hole 17 of the combination detection card can be independent of each other (i.e., isolated from each other), and the sample loading operation is performed separately; the first sample loading hole 16 and the second sample loading hole 17 can also be connected, and the sample loading operation is completed at one time.

[0091] The above-mentioned combined detection card is used in conjunction with a combined detection device for immune detection and cell analysis to achieve simultaneous detection of immune detection and cell analysis. The combined detection device includes at least an immune detection unit and a cell detection and analysis unit. The immune detection unit corresponds to the immune detection area formed by the immune detection component 300 on the combined detection card and is used to detect immune response signals; the cell detection and analysis unit corresponds to the detection surface formed by the cell analysis component on the combined detection card and is used to perform overall cell counting and / or cell typing analysis. This combined detection device can be modified from existing immune detection instruments and cell detection and analysis instruments. This combined detection device is not the focus of this application and will not be described in detail here.

[0092] The structural design of the embodiment 1 of the cell analysis component 400 is suitable for applications where the number of cells per unit volume is small or the abundance of the cell type is low, or where only cell counting or cell typing is required. According to the application, a test area 4 with a single thickness is selected and an appropriate thickness is set. , in order to perform high-precision measurement of cell number or high-precision analysis of single cell types.

[0093] Example 2

[0094] Figure 2A-2C The second embodiment of the cell analysis component is shown. The structure of the second embodiment is a further improvement on the structure of the first embodiment, and the difference between the second embodiment and the first embodiment is:

[0095] In order to further prevent the generation of bubbles, at least one exhaust hole 7 is provided on the test cavity 2. The exhaust hole 7 is a through hole connecting the interior of the test cavity 2 with the outside atmosphere. The exhaust hole 7 can be located on one side or both sides of the test area 4 or the drainage area 6. It can be a symmetrical or asymmetrical through hole, that is, the exhaust hole 7 passes through the side wall of one side of the test area 4 or the drainage area 6 or symmetrically passes through the two side walls of the test area 4 or the drainage area 6. Preferably, the exhaust hole 7 is a symmetrical inverted cone-shaped through hole, that is, the small end of the cone opens to the interior of the drainage area 6, and the large end of the cone opens to the outside atmosphere (see Figure 2C ).

[0096] The advantages of the inverted cone exhaust hole are that, first, it utilizes the surface tension between the liquid sample and the gas to make it easier to discharge bubbles. The exhaust method based on the inverted cone hole proposed in the present invention is not affected by the sampling angle and distance, and can effectively discharge bubbles. When the reagent needs to be pre-packaged in the test cavity 2, since the contact area between the reagent and the external environment is very small, when the reagent is added to the test cavity 2, its drying process is long. The design of the exhaust hole 7 can increase the contact area between the reagent and the external environment, accelerate the drying and uniform distribution of the reagent, thereby avoiding the generation of bubbles in many situations; it helps to accurately control the sample amount. After the liquid sample enters the exhaust hole 7, it is not easy to overflow due to the effect of surface tension; the exhaust hole 7 is set on one side or both sides of the test cavity 2. After the sampling is completed, only the side of the test cavity 2 needs to be wiped to avoid the loss of liquid sample due to wiping the injection port; at the same time, the inverted cone hole structure further reduces the possibility of sample loss by wiping.

[0097] Specifically, such as Figure 2A and Figure 2C As shown, there are two exhaust holes 7, namely a first exhaust hole 71 and a second exhaust hole 72, which are respectively located at the front end (the position before the liquid sample enters the test area 4) and the rear end (the position after the liquid sample flows out of the test area 4) of the drainage area 6, and are symmetrical inverted cone-shaped through holes that pass through the drainage area 6, which can further avoid the generation of bubbles.

[0098] The other structures of the second embodiment are the same as those of the first embodiment. For technical solutions not mentioned in the second embodiment, please refer to the first embodiment and will not be repeated here.

[0099] Accordingly, the cell analysis component 400 of this embodiment is installed in the combination test card. Based on the first embodiment of the combination test card, the structure of the test card cover 100 and / or the test card bottom shell 200 needs to be further improved. That is, corresponding connecting holes 20 are opened on the test card cover 100 and / or the test card bottom shell 200 at the position corresponding to the exhaust hole 7 of the cell analysis component, so that bubbles in the liquid sample can escape through the exhaust hole 7 and the connecting hole 20.

[0100] Specifically, the cell analysis component in this embodiment is provided with two exhaust holes, and two communication holes are opened at corresponding positions of the detection card cover and / or the detection card bottom shell.

[0101] Example 3

[0102] Figures 3A-3D The structure of the cell analysis component 400 of the combined detection card of the present invention is shown in the third embodiment. The structure of the third embodiment is a further improvement on the structure of the second embodiment. The difference between the third embodiment and the second embodiment is that:

[0103] In this embodiment, there are two test areas 4 located in the test cavity 2, namely the first test area 41 and the second test area 42. The two test areas are independent of each other (set at a distance) and connected. The two test areas are connected by the drainage area 6. In this embodiment, the two test areas are both rectangular and have a thickness of 、 , wherein the thickness of the first test area 41 is The detection surface formed by the liquid sample on the side wall 3 of the cavity in the first test area 41 has a large depth of field and a large sample carrying capacity per unit area, which is suitable for the overall accurate measurement of the number of cells. The thickness of the first test area 41 is Preferably, the thickness of the second test area 42 is 90 μm-120 μm; The depth of field of the detection surface formed by the liquid sample on the side wall 3 of the cavity in the second test area 42 is small, and the sample spreading area per unit volume is large, which can be used for precise differentiation of cell types. The thickness of the second test area 42 is The preferred thickness is 60 μm to 90 μm. This embodiment can simultaneously take into account the precision measurement of cell counting and typing. The two test areas 4 are connected by a drainage area 6 of uniform thickness.

[0104] Specifically, in this embodiment, four exhaust holes 7 are provided, namely a first exhaust hole 71, a second exhaust hole 72, a third exhaust hole 73, and a fourth exhaust hole 74. The first exhaust hole 71 and the second exhaust hole 72 are respectively located at the front end (the position before the liquid sample enters the first test area 41) and the middle of the rear end (the position where the liquid sample flows out of the second test area 42) of the drainage area 6. The third exhaust hole 73 is arranged on the first test area 41, and the fourth exhaust hole 74 is arranged on the second test area 42. The exhaust holes 7 are all symmetrical inverted cone-shaped through holes that penetrate the drainage area 6 or the test area 4, which can further avoid the generation of bubbles.

[0105] Obviously, each test area can be provided with a corresponding sampling port, and each sampling port is provided with a sampling notch. For test areas with thicker thickness, a larger liquid sample carrying capacity per unit area is required. By adding corresponding sampling ports, the insufficient liquid sample amount caused by a single sampling port can be avoided.

[0106] Specifically, multiple drainage areas 6 can be set (interconnected drainage areas of different thicknesses are called different drainage areas), each drainage area 6 corresponds to the corresponding test area 4, so as to facilitate the controlled and rapid flow of the liquid sample into the corresponding test area 4. By setting drainage areas 6 and test areas 4 of different thicknesses, different liquid flow rates and liquid laminar flow characteristics can be adjusted. The thickness of the test area 4 is less than the thickness of all drainage areas 6; similarly, one drainage area 6 can be set (interconnected drainage areas of the same thickness are called the same drainage area), and multiple test areas can correspond to the same drainage area.

[0107] Obviously, multiple test areas 4 may also have the same thickness, which is smaller than the thickness of the drainage area 6 . Setting different test areas 4 helps to analyze the accuracy and consistency of the test results of each test area 4 .

[0108] like Figures 5A to 5D FIG. 1 is a schematic structural diagram of a second embodiment of a combination test card according to the present invention. The cell analysis assembly 400 installed in the structure of the combination test card is also provided with an exhaust hole 7. Further improvements are required to the structure of the test card upper cover 100 and / or the test card bottom shell 200. Specifically, corresponding communication holes 20 are provided on the test card upper cover 100 and / or the test card bottom shell 200 at positions corresponding to the exhaust holes 7 of the cell analysis assembly, so that bubbles in the liquid sample can escape through the exhaust holes 7 and the communication holes 20.

[0109] Specifically, the cell analysis component 400 in this embodiment is provided with four exhaust holes, namely, a first exhaust hole 71, a second exhaust hole 72, a third exhaust hole 73, and a fourth exhaust hole 74. Then, four connecting holes are provided at corresponding positions on the test card cover 100 and / or the test card bottom shell 200, namely, the first connecting hole 201, the second connecting hole 202, the third connecting hole 203, and the fourth connecting hole 204 (the figure takes the example of the connecting holes provided on the test card cover 100).

[0110] The other structures of the cell analysis component 400 in the third embodiment are the same as those in the second embodiment. For technical solutions not mentioned in the third embodiment, please refer to the second or first embodiment, and will not be repeated here.

[0111] In the above embodiments, the material of the cell analysis component 400 of the present invention can be selected from any one or more combinations of optical-grade transparent polymers, glass, and quartz to ensure high light transmittance and low spontaneous fluorescence.

[0112] Obviously, the structure of the combined detection card of the present invention is not limited to the structure described in the above embodiment. On the basis of the concept of the present invention, simply increasing or decreasing the number of immune detection components 300 and cell analysis components 400, or changing their positions, shapes or combinations, all fall within the scope of the concept of the present invention.

[0113] Likewise, the structure of the cell analysis assembly 400 of the present invention is not limited to the structure described in the above embodiment. Based on the concept of the present invention, simply increasing or decreasing the number, changing the shape, position or combination of the test area, drainage area, injection port (with injection notch), exhaust hole and clamping part all fall within the scope of the concept of the present invention.

[0114] Combined detection method

[0115] Based on the above-mentioned combined detection card for immunoassay and cell analysis, the present invention also provides a combined detection method for immunoassay and cell analysis, and the specific operation steps are as follows:

[0116] S1) adding a liquid sample to the combination test card;

[0117] Specifically, use a pipette to absorb the original liquid sample (such as fresh urine, blood and other body fluids) or the pretreated liquid sample (the liquid after the original liquid sample is mixed with the reagent), and place the pipette at the first sample loading hole 16 and the second sample loading hole 17 of the combination detection card respectively to load the sample, so that the liquid sample flows into the sample pad 11 of the immunoassay component 300 and the sampling port 5 of the cell analysis component 400 respectively; or, use the pipette to add the liquid sample to the connected first sample loading hole 16 and the second sample loading hole 17 at one time, so that the liquid sample flows into the sample pad 11 of the immunoassay component 300 and the sampling port 5 of the cell analysis component 400 respectively.

[0118] S2) waiting for a predetermined time until an immune detection area is formed on the analysis membrane 13 of the immune detection component 300 and a detection surface is formed on the test area 4 of the cell analysis component 400;

[0119] Specifically, within a predetermined time, the liquid sample flows through the sample pad 11 of the immune detection component 300, and flows into the binding pad 12, the analysis membrane 13, and the absorbent paper 14 in sequence. During this process, a certain immune reaction occurs, and finally an immune detection area is formed on the analysis membrane 13; the liquid sample flows into the test area 4 through the injection port 5 and the drainage area 6 of the cell analysis component 400 and fills it, and finally a detection surface is formed in the test area 4.

[0120] S3) cooperating with the immune detection unit and the cell detection and analysis unit of the combined immune detection and cell analysis device to perform immune detection on the immune detection area of ​​the immune detection component of the combined detection card and to perform cell analysis on the detection surface of the cell analysis component;

[0121] Specifically, the combined detection card is placed in a predetermined position of the combined detection device for immune detection and cell analysis, and the immune detection unit collects the signal of the immune detection area of ​​the immune detection component 300 through the first interpretation window 18 and processes and analyzes it; the cell detection and analysis unit collects the signal of the detection surface of the cell analysis component 400 through the second interpretation window 19 and the third interpretation window 23 and processes and analyzes it.

[0122] The combined detection card of the above embodiment has the following technical effects:

[0123] A. The combined test card of the present invention integrates the immune detection component and the cell analysis component through the shell structure formed by the test card bottom shell and the test card top cover. When used in conjunction with a joint detection device equipped with an immune detection unit and a cell detection and analysis unit, it can simultaneously realize the immune detection and cell analysis functions, providing conditions for realizing synchronous on-site joint detection of immune and cell at the POCT level.

[0124] B. The test chamber 2 of the cell analysis assembly 400 of the present invention adopts a semi-open cavity structure. When reagent pre-packaging is required, the unobstructed air path accelerates the speed and uniformity of reagent packaging and prevents the formation of bubbles due to turbulent liquid flow during the subsequent injection process.

[0125] C. The cell analysis assembly 400 is provided with multiple test areas 4 of varying thicknesses. Thicker test areas 4 have a larger sample carrying capacity per unit area on the test surface, making them suitable for accurate overall cell number measurement. Thinner test areas 4 have a larger liquid sample spreading area per unit volume on the test surface, making them suitable for precise differentiation of cell types. This allows for accurate measurement of multiple cell parameters using the same liquid sample in a single test.

[0126] D. The shape, position (in the drainage area 6 and / or the test area 4), and number of the exhaust holes 7 can be designed to effectively exhaust gas and prevent bubbles while preventing the liquid sample from overflowing due to capillary flow inertia.

[0127] E. By setting a transition fillet on the inner edge of the cavity side wall 3 near the opening edge, the liquid at the end edge of the test cavity 2 can form a stable liquid bridge surface 10 to prevent the liquid sample from flowing out due to gravity during the movement of the combined test card.

[0128] Those skilled in the art should understand that these embodiments are only used to illustrate the present invention and do not limit the scope of the present invention. Various equivalent variations and modifications made to the present invention belong to the disclosure of the present invention.

Claims

1. A combined test card for immunoassay and cell analysis, characterized in that: include: A detection card upper cover (100), a detection card bottom shell (200), and an immunoassay component (300) and a cell analysis component (400) installed in a housing formed by the detection card upper cover (100) and the detection card bottom shell (200), wherein: The detection card bottom shell (200) is provided with a first placement slot (21) for installing the immune detection component (300), a second placement slot (22) for installing the cell analysis component (400), and a third interpretation window (23) for viewing the detection surface of the cell analysis component (400); The detection card upper cover (100) is provided with a first sample loading hole (16) for loading the sample of the immune detection component (300), a second sample loading hole (17) for loading the sample of the cell analysis component (400), a first interpretation window (18) for viewing the immune detection area of ​​the immune detection component (300), and a second interpretation window (19) for viewing the detection surface of the cell analysis component (400). The first interpretation window (18) is a window hole penetrating the detection card upper cover (100), the second interpretation window (19) and the third interpretation window (23) are window holes penetrating the detection card upper cover (100) and the detection card bottom shell (200) respectively, and the second interpretation window (19) and the third interpretation window (23) are aligned; The cell analysis component (400) comprises an assembly handle (1) and a test cavity (2) provided on the assembly handle (1). The test cavity (2) is a semi-open cavity formed by two parallel cavity side walls (3) with a certain gap, and has a test area (4), an injection port (5), and a drainage area (6) connecting the injection port (5) and the test area (4). The thickness of the test area (4) is Smaller than the thickness of the drainage area (6) , the thickness of the test area (4) The thickness of the drainage area (6) ranges from 60 μm to 120 μm. The range of the angle of the inlet (5) is 120 μm-500 μm, the inlet (5) is aligned with the second sample addition hole (17), and the test area (4) is aligned with the second interpretation window (19) and the third interpretation window (23); the inlet (5) is concave arc-shaped, and the angle α between the tangent of the downward arc and the horizontal reference plane of the inlet (5) is in the range of 15°-45°. The inlet (5) is located at the upper edge opening of the two cavity side walls (3) of the test cavity (2), wherein the upper edge of one of the two cavity side walls (3) located at the inlet (5) is provided with an inlet notch (8) so as to inject the liquid sample through the inlet notch (8); at least one exhaust hole (7) is provided on the test cavity (2), and the exhaust hole (7) is an inverted conical through hole, the small end of the cone opening is toward the inside of the test area (4) or the drainage area (6), and the large end of the cone opening is toward the outside of the test cavity (2).

2. The combined immunoassay and cell analysis test card according to claim 1, characterized in that: The first sample addition hole (16) and the second sample addition hole (17) are connected.

3. The combined test card for immunoassay and cell analysis according to claim 1, characterized in that: The exhaust hole (7) is a through hole connecting the interior of the test cavity (2) with the outside atmosphere, and penetrates one cavity side wall of the test area (4) or the drainage area (6) or symmetrically penetrates two cavity side walls (3) of the test area (4) or the drainage area (6); the exhaust hole (7) is connected to the outside atmosphere through a connecting hole (20) provided on the test card upper cover (100) and / or the test card bottom shell (200).

4. The combined immunoassay and cell analysis test card according to claim 1, wherein: One or more test areas (4) are provided in the test cavity (2), and the multiple test areas (4) are independent and connected to each other, and have the same or different thicknesses. The thickness of any test area (4) is smaller than the thickness of the drainage area (6). .

5. The combined test card for immunoassay and cell analysis according to claim 4, characterized in that: The test cavity (2) is provided with two test areas (4) of different thicknesses, namely a first test area (41) and a second test area (42), and the first test area (41) and the second test area (42) are connected by a drainage area (6).

6. The combined test card for immunoassay and cell analysis according to any one of claims 1 to 5, characterized in that: The injection notch (8) faces the second injection hole (17).

7. The combined test card for immunoassay and cell analysis according to any one of claims 1 to 5, characterized in that: The inner edge of the end edge of the cavity side wall (3) is provided with a transition fillet (10), and the R value of the transition fillet (10) ranges from 0.2 mm to 1.5 mm.

8. The combined test card for immunoassay and cell analysis according to any one of claims 1 to 5, characterized in that: The immunoassay component (300) is an immunochromatographic test paper, comprising a sample pad (11), a conjugation pad (12), an analysis membrane (13), absorbent paper (14) and an adhesive backing (15). The sample pad (11), the conjugation pad (12), the analysis membrane (13) and the absorbent paper (14) are sequentially adhered and fixed to the adhesive backing (15) in an overlapping manner. The sample pad (11) is aligned with the first sample addition hole (16), and the analysis membrane (13) is aligned with the first reading window (18).

9. A combined detection method using the combined detection card for immunoassay and cell analysis according to any one of claims 1 to 5, comprising the following steps: S1) adding a liquid sample to the sample well of the combination test card; S2) waiting for a predetermined time until an immune detection area is formed on the analysis membrane (13) of the immune detection component (300) and a detection surface is formed on the test area (4) of the cell analysis component (400); S3) Using an immunoassay instrument and a cell analysis instrument, respectively, to perform immunoassay on the immunoassay area of ​​the immunoassay component of the combined test card and to perform cell analysis on the detection surface of the cell analysis component.

10. The joint detection method according to claim 9, wherein: The sample addition operation in step S1) is as follows: Aspirate the liquid sample with a pipette, place the pipette at the first sample loading hole (16) and the second sample loading hole (17) of the combined detection card, and add the sample, so that the liquid sample flows into the sample pad (11) of the immunoassay component (300) and the sample inlet (5) of the cell analysis component (400), respectively; or The liquid sample is added to the connected first sample addition well (16) or the second sample addition well (17) at one time through the pipette, so that the liquid sample flows into the sample pad (11) of the immunoassay component (300) and the sample inlet (5) of the cell analysis component (400) respectively.

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