Blood type rapid detection immunochromatography reagent strip and preparation method thereof

Abstract

The invention discloses a blood type rapid detection immunochromatography reagent strip and a preparation method thereof. The blood type rapid detection immunochromatography reagent strip comprises a test strip, the test strip comprises a bottom plate, a sample pad, a nitrocellulose membrane and water absorption paper, the sample pad, the nitrocellulose membrane and the water absorption paper are sequentially arranged on the bottom plate in an overlapped mode, and the sample pad, the nitrocellulose membrane and the water absorption paper are sequentially arranged on the bottom plate in an overlapped mode. The nitrocellulose membrane is provided with a first detection line, a second detection line and a quality control line which are arranged at intervals, the first detection line is coated with an anti-A-type blood antibody, the second detection line is coated with an anti-B-type blood antibody, and the quality control line is coated with an anti-erythrocyte antibody. The blood type rapid detection immunochromatography reagent strip can achieve rapid detection of blood types and is high in specificity, simple in operation step, extremely small in needed sample size, high in detection speed and visual in detection result.

Description

Technical Field

[0001] The invention relates to the technical field of biological detection, and in particular to a blood type rapid detection immunochromatographic reagent strip and a preparation method thereof. Background Art

[0002] For blood type detection, the existing technologies mainly include molecular biology technology, blood type identification technology based on sequencing reaction and serological methods. Among them, molecular biology technology mainly includes the following technologies: (1) Restriction fragment length polymorphism PCR technology, which detects relevant blood type gene fragments. Since restriction endonucleases only target specific nucleic acid cleavage sites, it has high specificity for red blood cell blood type identification. The disadvantage is that it takes a long time, at least 6 hours, and is limited by the restriction site, so it is not suitable for all gene polymorphism detection. At the same time, the use of endonucleases also increases the detection cost. (2) Sequence-specific primer PCR technology, this technology does not require endonuclease reaction for red blood cell blood type allele detection, simplifies the operation procedure, saves detection costs, and the detection time depends more on the length of the amplified red blood cell blood type allele fragment, but it is not as specific as restriction fragment length polymorphism PCR technology in judging homozygotes and heterozygotes. (3) Multiplex PCR technology. Although multiplex PCR technology has the advantage of detection throughput, it also has the problem of requiring the annealing, extension and denaturation temperatures of multiple pairs of primers to be relatively consistent, which limits the detection throughput. The false positive rate or false negative rate is high. Multiple PCR products make it difficult to judge the results during electrophoresis analysis. The sensitivity of the analytical instrument is also required to be higher. When designing probe primers, it is necessary to add non-specific oligonucleotide fragments of different lengths to distinguish the length of product fragments. The operation procedure is complicated and time-consuming. Therefore, multiplex PCR technology has not achieved high-throughput detection in the true sense. (4) Timely quantitative PCR technology. Compared with restriction enzyme fragment length polymorphism PCR technology and sequence-specific primer PCR technology, this technology has obvious advantages in the detection time of red blood cell blood type identification. The procedure is simpler and can be quantitatively detected in time, but the detection throughput is not as high as that of multiplex PCR technology. (5) PCR-enzyme-linked immunosorbent assay technology. This technology is complicated and time-consuming.

[0003] Blood typing technologies based on sequencing reactions mainly include Sanger sequencing technology, massively parallel sequencing technology, pyrophosphate sequencing technology, and Ion Torrent sequencing technology, but they have low detection speed and lack advantages in detection throughput.

[0004] Serological methods mainly include the following technologies: (1) Slide method. Slide method is a direct hemagglutination test. The serum and the blood sample to be tested are dripped onto a slide. The red blood cell type is determined by observing the agglutination of red blood cells. The advantage of this method for identifying red blood cell types is that it is simple and fast to operate. However, the sensitivity of this method is poor. It can only be used for the initial detection of red blood cell types. The final identification of red blood cell types requires the use of other methods or re-testing. (2) Test tube method. The hemagglutination test of the test tube method is carried out in a test tube. The operation is not as simple as the slide method. The blood sample needs to be centrifuged. For the agglutinated blood samples that are difficult to identify, microscopic observation is required. When the test tube method uses saline agglutination to identify red blood cell types, non-specific interference is prone to occur, resulting in inconsistency between positive and negative typing. (3) Microcolumn gel method: The microcolumn gel method determines the agglutination of the blood sample by observing whether the blood sample can pass through the molecular sieve composed of gel particles, and then identifies the red blood cell type. This method requires less sample, has a high degree of automation, and has higher sensitivity and specificity. However, in terms of operation, the results of this method are easy to judge and easy to preserve, but the detection time is relatively long and it is not suitable for emergency identification of red blood cell types.

[0005] It can be seen that the molecular biology methods and gene sequencing methods in the existing technology have long detection time and high cost, and have limitations in situations where blood transfusion is urgently needed. The slide method of serological methods has poor sensitivity, the test tube method is relatively difficult to operate, and the microcolumn gel method is relatively time-consuming, and is not suitable for emergency identification of red blood cell types. Summary of the invention

[0006] Based on this, it is necessary to provide a blood type rapid detection immunochromatographic reagent strip. The blood type rapid detection immunochromatographic reagent strip of the present invention can realize rapid detection of blood type, has strong specificity, simple operation steps, requires very little sample volume, has fast detection speed, and has intuitive detection results.

[0007] An embodiment of the present application provides an immunochromatographic reagent strip for rapid blood type detection.

[0008] A blood type rapid detection immunochromatographic reagent strip comprises a test strip, wherein the test strip comprises a bottom plate, a sample pad, a nitrocellulose membrane and blotting paper, wherein the sample pad, the nitrocellulose membrane and the blotting paper are sequentially overlapped and arranged on the bottom plate, a first detection line, a second detection line and a quality control line which are arranged at intervals are arranged on the nitrocellulose membrane, the first detection line is coated with anti-type A blood antibodies, the second detection line is coated with anti-type B blood antibodies, and the quality control line is coated with anti-red blood cell antibodies.

[0009] In some embodiments, the first detection line, the second detection line, and the quality control line on the nitrocellulose membrane are arranged in sequence, wherein the first detection line is closest to the sample pad.

[0010] In some embodiments, the bottom plate is in a long strip structure, and the sample pad, the nitrocellulose membrane and the absorbent paper all extend along the length direction of the bottom plate.

[0011] In some of the embodiments, the base plate is a P VC plate.

[0012] In some embodiments, the blood type rapid detection immunochromatographic reagent strip also includes a shell, on which is provided a detection window, a sample loading hole and a flushing hole, the test strip is assembled in the shell, the first detection line, the second detection line and the quality control line on the nitrocellulose membrane are located correspondingly at the detection window position, the sample loading hole corresponds to the nitrocellulose membrane, and the flushing hole corresponds to the sample pad.

[0013] In some embodiments, the size of the flushing hole gradually decreases from a side located on the outer surface of the shell to a side located inside the shell.

[0014] In some embodiments, the detection window is in the shape of an elongated strip, and the first detection line, the second detection line, and the quality control line are all located in the middle of the detection window.

[0015] An embodiment of the present application also provides a method for preparing a blood type rapid detection immunochromatographic reagent strip.

[0016] A method for preparing a blood type rapid detection immunochromatographic reagent strip comprises the following steps:

[0017] Prepare a first test line working solution, a second test line working solution and a quality control line working solution;

[0018] Using the first detection line working solution, the second detection line working solution and the quality control line working solution to respectively draw a first detection line, a second detection line and a quality control line on the nitrocellulose membrane, and placing the nitrocellulose membrane in a drying oven at 50° C. to 60° C. to dry for 24 h to 48 h;

[0019] The sample pad, nitrocellulose membrane and absorbent paper are assembled and connected to the bottom plate, and the assembled intermediate product is cut into test strips of a predetermined width.

[0020] In some embodiments, the preparation method includes at least one of the following conditions:

[0021] (1) diluting the anti-blood type A antibody to 0.5 mg / mL to 1 mg / mL using 0.01 M Tris buffer at pH 8 containing 3% to 5% trehalose to obtain the first test line working solution;

[0022] (2) diluting the anti-type B blood antibody to 0.1 mg / mL to 0.2 mg / mL using 0.01 M Tris buffer at pH 8 containing 3% to 5% trehalose to obtain the second test line working solution;

[0023] (3) Dilute the anti-erythrocyte antibody to 0.01 mg / mL to 0.05 mg / mL using 0.01 M Tris buffer at pH 8 containing 3% to 5% trehalose to obtain the quality control line working solution.

[0024] In some embodiments, the preparation method includes at least one of the following conditions:

[0025] (1) The first detection line drawn on the nitrocellulose membrane has a drawing volume of 0.2 μL / cm to 2 μL / cm;

[0026] (2) The amount of the second detection line drawn on the nitrocellulose membrane is 0.2 μL / cm~2 μL / cm;

[0027] (3) The amount of the quality control line drawn on the nitrocellulose membrane is 0.2 μL / cm~2 μL / cm.

[0028] In some embodiments, the preparation method includes at least one of the following conditions:

[0029] (1) Cutting the dried nitrocellulose membrane into long strips, wherein the length of the nitrocellulose membrane is 30 cm to 40 cm, and the width of the nitrocellulose membrane is 2.2 cm to 3 cm;

[0030] (2) The length of the absorbent paper is 30 cm to 40 cm, and the width of the absorbent paper is 1.5 cm to 2 cm;

[0031] (3) Install the assembled test strip in a housing having a detection window, a sample loading hole, and a flushing hole, wherein the first detection line, the second detection line, and the quality control line on the nitrocellulose membrane are located at the detection window position, the sample loading hole corresponds to the nitrocellulose membrane, and the flushing hole corresponds to the sample pad.

[0032] The above-mentioned blood type rapid detection immunochromatographic reagent strip can realize rapid detection of blood type, with simple operation steps, very small sample volume, fast detection speed, and results in 30s~3min. No instrument is required to assist in the test, and blood type can be quickly known when blood transfusion is urgently needed in the clinic, providing more time for saving lives. The above-mentioned blood type rapid detection immunochromatographic reagent strip has intuitive detection results. By observing whether a red line appears at the corresponding position, the corresponding blood type can be known. No colorimetric card comparison is required. The detection results are more intuitive than traditional slide methods and chromatography methods. Users can quickly and clearly know the tested blood type, which greatly facilitates the clinical application needs of hospitals and other medical places. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings based on these drawings without creative work.

[0034] In order to more completely understand the present application and its beneficial effects, the following description will be given in conjunction with the accompanying drawings. In the following description, the same reference numerals represent the same parts.

[0035] FIG1 is a schematic diagram of a blood type rapid detection immunochromatographic reagent strip according to an embodiment of the present invention;

[0036] FIG2 is a schematic diagram of a test strip of a blood type rapid detection immunochromatographic reagent strip according to an embodiment of the present invention;

[0037] FIG3 is a schematic top view of a test strip of a blood type rapid detection immunochromatographic reagent strip according to an embodiment of the present invention;

[0038] FIG4 is a schematic diagram of an immunochromatographic reagent strip for rapid detection of blood type according to another embodiment of the present invention.

[0039] Description of Reference Numerals

[0040] 10. Blood type rapid detection immunochromatographic reagent strip; 100. Test strip; 101. Bottom plate; 102. Sample pad; 103. Nitrocellulose membrane; 104. Absorbent paper; 105. First detection line; 106. Second detection line; 107. Quality control line; 200. Shell; 201. Shell base; 202. Shell cover; 2031. Sample loading hole; 2032. Flushing hole; 204. Detection window; 2041. First detection sub-window; 2042. Second detection sub-window; 205. Marking area; 206. Installation station; 207. Limiting piece; 208. Baffle; 209. Card slot; 210. Buckle. DETAILED DESCRIPTION

[0041] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below in conjunction with the accompanying drawings. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific embodiments disclosed below.

[0042] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as limiting the present invention.

[0043] In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0044] In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may mean that the first and second features are in direct contact, or the first and second features are in indirect contact through an intermediate medium. Moreover, a first feature being "above", "above" or "above" a second feature may mean that the first feature is directly above or obliquely above the second feature, or simply means that the first feature is higher in level than the second feature. A first feature being "below", "below" or "below" a second feature may mean that the first feature is directly below or obliquely below the second feature, or simply means that the first feature is lower in level than the second feature.

[0045] In the description of the present invention, "several" means more than one, "many" means more than two, "greater than", "less than", "exceed", etc. are understood to exclude the number itself, and "above", "below", "within", etc. are understood to include the number itself. If there is a description of "first" or "second", it is only used for the purpose of distinguishing the technical features, and cannot be understood as indicating or implying the relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0046] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0047] In this article, "optionally", "optional", "optional" means optional, that is, it means to be selected from any of the two parallel solutions of "yes" or "no". If there are multiple "optional" in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "optional" is independent. In this application, the descriptions such as "optionally contain" and "optionally include" mean "contain or not contain".

[0048] In the present application, when it comes to a numerical interval (i.e., a numerical range), unless otherwise specified, the distribution of the optional numerical values ​​in the numerical interval is considered to be continuous, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical interval, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to an integer in the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints, which is equivalent to directly listing each integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be combined. In other words, unless otherwise specified, the numerical range disclosed in the present application should be understood to include any and all sub-ranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. "Numerical interval" allows for a broad range of quantitative intervals such as percentage intervals, ratio intervals, and ratio intervals.

[0049] The embodiment of the present application provides a blood type rapid detection immunochromatographic reagent strip 10 to solve the problem that the molecular biology method and gene sequencing method have long detection time and high cost in the existing prior art for blood type detection, and have limitations in the case of emergency blood transfusion; the serological method has the problem that the operation is relatively difficult and the detection time is relatively long, and it is not suitable for emergency identification of red blood cell blood types. The blood type rapid detection immunochromatographic reagent strip 10 will be described below in conjunction with the accompanying drawings.

[0050] The blood type rapid detection immunochromatographic reagent strip 10 provided in the embodiment of the present application is exemplified in FIG1 , which is a schematic diagram of the structure of the blood type rapid detection immunochromatographic reagent strip 10 provided in the embodiment of the present application. The blood type rapid detection immunochromatographic reagent strip 10 of the present application can be used for rapid, high-throughput detection of blood types.

[0051] In order to more clearly illustrate the structure of the blood typing rapid detection immunochromatographic reagent strip 10 , the blood typing rapid detection immunochromatographic reagent strip 10 will be introduced below with reference to the accompanying drawings.

[0052] Exemplarily, please refer to FIG. 1 and FIG. 2 , a blood type rapid detection immunochromatographic reagent strip 10 includes a test strip 100. Please refer to FIG. 2 , which is a schematic diagram of a test strip of a blood type rapid detection immunochromatographic reagent strip according to an embodiment of the present invention, and the test strip 100 includes a base plate 101, a sample pad 102, a nitrocellulose membrane 103 and a blotting paper 104. Among them, the sample pad 102, the nitrocellulose membrane 103 and the blotting paper 104 are overlapped and arranged on the base plate 101 in sequence. The nitrocellulose membrane 103 is provided with a first detection line 105, a second detection line 106 and a quality control line 107 which are arranged at intervals. The first detection line 105 is coated with anti-type A blood antibodies. The second detection line 106 is coated with anti-type B blood antibodies. The quality control line 107 is coated with anti-red blood cell antibodies.

[0053] In some embodiments, the base plate 101 is a PVC board. PVC board is a polyvinyl chloride board, which is a material widely used in the fields of construction, furniture manufacturing, advertising, decoration, etc. PVC board has many advantages, making it a preferred material in many applications. In this application, the advantages of using PVC board for the base plate 101 include (1) Weather resistance: PVC board has good resistance to ultraviolet rays and climate change, and is suitable for outdoor use. (2) Waterproof and moisture-proof: PVC material does not absorb water and is suitable for storage in a humid environment. (3) Corrosion resistance: It has good resistance to acids, alkalis and most chemicals. (4) Lightweight: Compared with metal or wooden boards, PVC boards are lighter and easier to carry and install. (5) Easy to process: It can be easily cut, drilled, bent and welded, which is convenient for customization and design. (6) Cost-effectiveness: Compared with other materials, PVC boards are usually cheaper and more cost-effective.

[0054] In some embodiments, referring to FIG. 1 and FIG. 2 , the first detection line 105 , the second detection line 106 and the quality control line 107 on the nitrocellulose membrane 103 are sequentially arranged, wherein the first detection line 105 is closest to the sample pad 102 .

[0055] In some of the embodiments, please refer to Figures 2 and 3, Figure 3 is a top view of a test strip of a blood type rapid detection immunochromatographic reagent strip according to an embodiment of the present invention, the bottom plate 101 is a long strip structure, and the sample pad 102, nitrocellulose membrane 103 and absorbent paper 104 all extend along the length direction of the bottom plate 101.

[0056] In some embodiments, please refer to FIG. 1 , the blood type rapid detection immunochromatographic reagent strip 10 further includes a housing 200. The housing 200 is provided with a detection window 204, a sample loading hole 2031, and a flushing hole 2032. The test strip 100 is assembled in the housing 200. The first detection line 105, the second detection line 106, and the quality control line 107 on the nitrocellulose membrane 103 are located at the detection window 204, the sample pad 102 is located at the flushing hole 2032, and the sample loading hole 3021 corresponds to the nitrocellulose membrane 103.

[0057] In some embodiments, the size of the flushing hole 2032 is gradually reduced from one side located on the outer surface of the housing 200 to one side located inside the housing 200. In the present application, the size of the flushing hole 2032 is gradually reduced from one side located on the outer surface of the housing 200 to one side located inside the housing 200, which is convenient for adding samples and for flushing liquid to enter the flushing hole 2032.

[0058] In some embodiments, the detection window 204 is in the shape of a long strip. The first detection line 105 , the second detection line 106 and the quality control line 107 are all located in the middle of the detection window 204 .

[0059] In some embodiments, the housing 200 includes a housing base 201 and a housing cover 202, which can be assembled with each other and can accommodate the test strip 100. The housing cover 202 is provided with the above-mentioned detection window 204, sample addition hole 2031, and flushing hole 2032.

[0060] In some embodiments, the outer surface of the housing cover 202 is further provided with a logo area 205. The logo area 205 may be provided with a label, a company trademark, a one-dimensional code or a two-dimensional code of a product, and the like.

[0061] In some embodiments, as shown in FIG1 , a mounting station 206 is disposed on the housing base 201. The mounting station 206 is a long strip structure, and the mounting station 206 is used to accommodate the test strip 100.

[0062] Preferably, as shown in FIG1 , a limiter 207 is provided on the shell base 201. The two ends of the installation station 206 in the length direction are respectively provided with limiters 207, which have a certain height. The limiters 207 can limit the test strip 100 on one hand, and can support the shell cover 202 on the other hand.

[0063] It should be noted that, as shown in FIG. 1 , the installation station 206 may be in the tank body, or may be an area surrounded by positioning strips.

[0064] In some embodiments, as shown in FIG1 , the limiting member 207 is in a U-shaped structure as a whole. The limiting member 207 of the U-shaped structure can limit the end of the test strip 100 .

[0065] In some embodiments, as shown in FIG1 , at least one baffle 208 is disposed on both sides of the installation station 206. The baffle 208 is used to limit the position of the test strip 100. The structures and shapes of different baffles 208 may be the same or different.

[0066] In some of the embodiments, please refer to FIG. 1 , a slot 209 and a buckle 210 are respectively provided on the shell base 201 and the shell cover 202 , and the slot 209 and the buckle 210 can be embedded and matched with each other to realize a detachable connection between the shell base 201 and the shell cover 202 .

[0067] Preferably, referring to FIG. 1 , the card slot 209 and the card buckle 210 are located at the corner positions on the shell base 201 and the shell cover 202 .

[0068] The above-mentioned blood type rapid detection immunochromatographic reagent strip 10 can realize rapid detection of blood type, with simple operation steps, very small sample volume, fast detection speed, and results in 30s~3min, without the need for instrument-assisted testing, and blood type can be quickly known when blood transfusion is urgently needed in the clinic, providing more time for saving lives. The detection result of the above-mentioned blood type rapid detection immunochromatographic reagent strip 10 is intuitive, and the corresponding blood type can be known by observing whether a red line appears at the corresponding position, without the need for colorimetric card comparison. The detection result is more intuitive than the traditional slide method and chromatography method, and the user can quickly and clearly know the tested blood type, which greatly facilitates the clinical application needs of medical places such as hospitals.

[0069] An embodiment of the present application also provides a method for preparing a blood type rapid detection immunochromatographic reagent strip 10 .

[0070] A method for preparing a blood type rapid detection immunochromatographic reagent strip 10 comprises the following steps:

[0071] Prepare the first test line working solution, the second test line working solution and the quality control line working solution.

[0072] The first detection line 105, the second detection line 106 and the quality control line 107 are respectively drawn on the nitrocellulose membrane 103 using the first detection line working solution, the second detection line working solution and the quality control line working solution, and the nitrocellulose membrane 103 is placed in a drying oven at 50°C to 60°C to dry for 24h to 48h.

[0073] The sample pad 102 , the nitrocellulose membrane 103 and the absorbent paper 104 are assembled and connected to the bottom plate 101 , and the assembled intermediate product is cut into test strips 100 of a predetermined width.

[0074] In some of the embodiments, the anti-blood type A antibody is diluted to 0.5 mg / mL to 1 mg / mL with 0.01 M Tris buffer at pH 8 containing 3 to 5% trehalose to obtain a first detection line working solution.

[0075] In some of the embodiments, the anti-type B blood antibody is diluted to 0.1 mg / mL-0.2 mg / mL with 0.01 M Tris buffer at pH 8 containing 3-5% trehalose to obtain the second detection line working solution.

[0076] In some of the embodiments, the anti-erythrocyte antibody is diluted to 0.01 mg / mL to 0.05 mg / mL with 0.01 M Tris buffer at pH 8 containing 3% to 5% trehalose to obtain a quality control line working solution.

[0077] In some embodiments, the amount of the first detection line 105 drawn on the nitrocellulose membrane 103 is 0.2 μL / cm~2 μL / cm. For example, the amount of the first detection line 105 drawn on the nitrocellulose membrane 103 is 0.2 μL / cm; for another example, the amount of the first detection line 105 drawn on the nitrocellulose membrane 103 is 2 μL / cm. It is not difficult to understand that in other examples, the amount of the first detection line 105 drawn on the nitrocellulose membrane 103 can also be 1.2 μL / cm, 1.5 μL / cm, 1.8 μL / cm or other values ​​and the range between any two of the above values.

[0078] In some embodiments, the amount of the second detection line 106 drawn on the nitrocellulose membrane 103 is 0.2 μL / cm~2 μL / cm. For example, the amount of the second detection line 106 drawn on the nitrocellulose membrane 103 is 0.2 μL / cm; for another example, the amount of the second detection line 106 drawn on the nitrocellulose membrane 103 is 2 μL / cm. It is not difficult to understand that in other examples, the amount of the second detection line 106 drawn on the nitrocellulose membrane 103 can also be 1.2 μL / cm, 1.5 μL / cm, 1.8 μL / cm or other values ​​and the range between any two of the above values.

[0079] In some embodiments, the amount of the quality control line 107 drawn on the nitrocellulose membrane 103 is 0.2 μL / cm~2 μL / cm. For example, the amount of the quality control line 107 drawn on the nitrocellulose membrane 103 is 0.2 μL / cm; for another example, the amount of the quality control line 107 drawn on the nitrocellulose membrane 103 is 2 μL / cm. It is not difficult to understand that in other examples, the amount of the quality control line 107 drawn on the nitrocellulose membrane 103 can also be 1.2 μL / cm, 1.5 μL / cm, 1.8 μL / cm or other values ​​and the range between any two of the above values.

[0080] In some embodiments, the dried nitrocellulose membrane 103 is cut into strips, the length of the nitrocellulose membrane 103 is 30 cm to 40 cm, and the width of the nitrocellulose membrane 103 is 2.2 cm to 3 cm. The length and width of the nitrocellulose membrane 103 can be set according to actual needs.

[0081] In some embodiments, the length of the absorbent paper 104 is 30 cm to 40 cm, and the width of the absorbent paper 104 is 1.5 cm to 2 cm.

[0082] In some of the embodiments, the assembled test strip 100 is installed in a housing 200 having a detection window 204, a sample loading hole 2031 and a flushing hole 2032, the first detection line 105, the second detection line 106 and the quality control line 107 on the nitrocellulose membrane 103 are located at the detection window 204, the sample loading hole 3021 corresponds to the nitrocellulose membrane 103, and the sample pad 102 is located at the flushing hole 2032.

[0083] In some embodiments, see FIG. 4, which is a schematic diagram of a blood type rapid detection immunochromatographic reagent strip according to another embodiment of the present invention. In FIG. 4, the detection window 204 includes a first detection sub-window 2041 and a second detection sub-window 2042, wherein the quality control line 107 corresponds to the first detection sub-window 2041, the second detection line 106 corresponds to the sample addition hole 3021, and the first detection line 105 corresponds to the second detection sub-window 2042. Samples can be added to the sample addition hole 3021. When the quality control line 107 and the first detection line 105 in the first detection sub-window 2041 and the second detection sub-window 2042 are colored, the detection sample is type A blood. When the quality control line 107 and the second detection line 106 in the first detection sub-window 2041 and the sample addition hole 3021 are colored, the detection sample is type B blood. When the first detection sub-window 2041, the second detection sub-window 2042, the quality control line 107, the first detection line 105, and the second detection line 106 in the sample addition hole 3021 are all colored, the detected sample is AB type blood. The detection principle of the structure shown in FIG1 is the same as above.

[0084] The blood type rapid detection immunochromatographic reagent strip 10 of the present application provides a blood type initial screening detection structure with high sensitivity, good repeatability, high specificity, fast detection speed, simple operation, intuitive detection results, and economical and practical through the combination of antigen and antibody, and realizes the qualitative detection of blood type A antigen and blood type B antigen in whole blood. For example, the source of whole blood includes samples collected by EDTA-K2 anticoagulant tube, heparin lithium anticoagulant tube, heparin sodium anticoagulant tube, sodium citrate anticoagulant tube, etc. The basic principle of the present application is to use the specific combination of antigen and antibody and the red mark of red blood cells themselves to coat anti-A blood type antibodies, anti-B blood type antibodies, and anti-red blood cell antibodies on the nitrocellulose membrane 103. When the whole blood contacts the antibodies coated on the nitrocellulose membrane 103, the specific antigens on the surface of red blood cells expressing the blood type bind to the antibodies, and the position of the corresponding blood type antibodies coated on the nitrocellulose membrane 103 shows a red strip. Red blood cells of all blood types will bind to the anti-red blood cell antibodies coated on the nitrocellulose membrane 103, so a red strip will appear at the location where the anti-red blood cell antibodies are coated on the nitrocellulose membrane 103. This strip serves as the quality control line 107 to determine whether the sample has been added. That is, when the quality control line 107 in the first detection sub-window 2041 does not show color, it indicates that the test is invalid and needs to be retested.

[0085] In summary, the blood type rapid detection immunochromatographic reagent strip 10 provided in the embodiment of the present application has the following beneficial effects compared with the prior art:

[0086] (1) The blood type rapid detection immunochromatographic reagent strip 10 of the present application has a simple structure and simple operation steps. The sample volume required for the test is extremely small. It only needs to take 10 μL of whole blood and add it to the diluent, mix it evenly, take 10 μL of the sample to be tested and add it to the sample adding hole 2031 of the housing 200, and then add 50 μL-80 μL of flushing liquid to the flushing hole 2032 for testing. The present application adds the sample to the sample adding hole 2031 and adds the flushing liquid to the flushing hole 2032, which can develop the color faster.

[0087] (2) The first test line 105, the second test line 106 and the quality control line 107 in the present application are on the same reagent strip, so the test result can be obtained by adding the sample only once, which simplifies the operation steps.

[0088] (3) The test is fast, with results available in 30 seconds to 3 minutes. No instrument is required to assist in the test. When blood transfusion is urgently needed, the blood type can be quickly determined, providing more time to save lives.

[0089] (4) The test results are intuitive. The corresponding blood type can be known by observing whether a red line appears at the corresponding position on the test card. No colorimetric card is required for comparison. Compared with the slide method and chromatography method in traditional technologies, the test results of this application are more intuitive, and the blood type of the corresponding line can be indicated on the card shell so that the user can know the blood type of the person being tested at a glance.

[0090] In the above embodiments, the description of each embodiment has its own emphasis. For parts that are not described in detail in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

[0091] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0092] The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A blood type rapid detection immunochromatographic reagent strip, characterized in that: The invention comprises a test strip, which comprises a bottom plate, a sample pad, a nitrocellulose membrane and blotting paper, wherein the sample pad, the nitrocellulose membrane and the blotting paper are overlapped in sequence on the bottom plate, a first detection line, a second detection line and a quality control line are arranged at intervals on the nitrocellulose membrane, the first detection line is coated with anti-type A blood antibodies, the second detection line is coated with anti-type B blood antibodies, and the quality control line is coated with anti-red blood cell antibodies.

2. The blood type rapid detection immunochromatographic reagent strip according to claim 1, characterized in that: The first detection line, the second detection line and the quality control line on the nitrocellulose membrane are arranged in sequence, wherein the first detection line is closest to the sample pad.

3. The blood type rapid detection immunochromatographic reagent strip according to claim 1, characterized in that: The blood type rapid detection immunochromatographic reagent strip meets at least one of the following conditions: (1) The base plate includes a P VC plate; (2) The bottom plate is in the shape of a long strip, and the sample pad, the nitrocellulose membrane and the absorbent paper all extend along the length direction of the bottom plate.

4. The blood type rapid detection immunochromatographic reagent strip according to any one of claims 1 to 3, characterized in that: The blood type rapid detection immunochromatographic reagent strip also includes a shell, on which a detection window, a sample addition hole and a flushing hole are provided. The test strip is assembled in the shell, and the first detection line, the second detection line and the quality control line on the nitrocellulose membrane are located correspondingly at the detection window position, the sample addition hole corresponds to the nitrocellulose membrane, and the flushing hole corresponds to the sample pad.

5. The blood type rapid detection immunochromatographic reagent strip according to claim 4, characterized in that: The size of the flushing hole gradually decreases from the side located on the outer surface of the shell to the side located inside the shell.

6. The blood type rapid detection immunochromatographic reagent strip according to claim 4, characterized in that: The detection window is in the shape of a long strip, and the first detection line, the second detection line and the quality control line are all located in the middle of the detection window.

7. A method for preparing a blood type rapid detection immunochromatographic reagent strip according to any one of claims 1 to 6, characterized in that: Including steps such as: Prepare a first test line working solution, a second test line working solution and a quality control line working solution; Using the first detection line working solution, the second detection line working solution and the quality control line working solution to respectively draw a first detection line, a second detection line and a quality control line on the nitrocellulose membrane, and placing the nitrocellulose membrane in a drying oven at 50° C. to 60° C. to dry for 24 h to 48 h; The sample pad, nitrocellulose membrane and absorbent paper are assembled and connected to the bottom plate, and the assembled intermediate product is cut into test strips of a predetermined width.

8. The method for preparing the blood type rapid detection immunochromatographic reagent strip according to claim 7, characterized in that: The preparation method comprises at least one of the following conditions: (1) diluting the anti-blood type A antibody to 0.5 mg / mL to 1 mg / mL using 0.01 M Tris buffer at pH 8 containing 3% to 5% trehalose to obtain the first test line working solution; (2) diluting the anti-type B blood antibody to 0.1 mg / mL to 0.2 mg / mL using 0.01 M Tris buffer at pH 8 containing 3% to 5% trehalose to obtain the second test line working solution; (3) Dilute the anti-erythrocyte antibody to 0.01 mg / mL to 0.05 mg / mL using 0.01 M Tris buffer at pH 8 containing 3% to 5% trehalose to obtain the quality control line working solution.

9. The method for preparing the blood type rapid detection immunochromatographic reagent strip according to claim 7 or 8, characterized in that: The preparation method comprises at least one of the following conditions: (1) The first detection line drawn on the nitrocellulose membrane has a drawing volume of 0.2 μL / cm to 2 μL / cm; (2) The amount of the second detection line drawn on the nitrocellulose membrane is 0.2 μL / cm~2 μL / cm; (3) The quality control line drawn on the nitrocellulose membrane has a draw volume of 0.2 μL / cm~2 μL / cm.

10. The method for preparing the blood type rapid detection immunochromatographic reagent strip according to claim 7 or 8, characterized in that: The preparation method comprises at least one of the following conditions: (1) Cutting the dried nitrocellulose membrane into long strips, wherein the length of the nitrocellulose membrane is 30 cm to 40 cm, and the width of the nitrocellulose membrane is 2.2 cm to 3 cm; (2) The length of the absorbent paper is 30 cm to 40 cm, and the width of the absorbent paper is 1.5 cm to 2 cm; (3) Install the assembled test strip in a housing having a detection window, a sample loading hole, and a flushing hole, wherein the first detection line, the second detection line, and the quality control line on the nitrocellulose membrane are located at the detection window position, the sample loading hole corresponds to the nitrocellulose membrane, and the flushing hole corresponds to the sample pad.

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