Erythrocyte blood type detection and antibody screening identification system and method

Abstract

The invention discloses a red blood cell blood type detection and antibody screening identification system and method. The system comprises a positive typing detection liquid phase exclusion chromatography separation medium, a reverse typing detection liquid phase exclusion chromatography separation medium, an antibody screening liquid phase exclusion chromatography separation medium and a micro-column card, the chromatographic separation media are respectively filled in the micro-column card, and are centrifuged to observe an observation window of the micro-column card so as to independently realize red blood cell blood type positive typing detection, red blood cell blood type negative typing detection and red blood cell antibody screening identification. According to the detection scheme, clinical erythrocyte blood group compatibility detection is promoted, various blood group systems such as ABO, Rh, MNS, Kell and Kidd of erythrocytes can be rapidly and accurately detected, different detection items can be selected according to POCT (point-of-care testing) requirements, and the method is suitable for various blood group detection conditions such as hospitals, families and field emergency.

Description

Technical Field

[0001] The present invention relates to the technical field of blood immune detection, and in particular to a system and method for red blood cell type detection and antibody screening and identification. Background Art

[0002] There are 45 blood type systems that have been discovered and recognized by the International Society of Blood Transfusion, among which the ABO and Rh blood type systems are of the most important clinical significance. It is mainly manifested in medical blood transfusions used for clinical diagnosis and treatment. If the transfused blood is incompatible with the patient, transfusion reactions will occur, causing fever, mild dyspnea and shock, and even death. Other blood type systems include MNS, Kidd, Kell, Duffy, etc. Incompatible blood type transfusions that trigger allogeneic immune reactions also threaten the life safety of patients. Blood typing methods include paper card method, test tube method, microcolumn gel method and microplate method. Both paper card method and test tube method are manually operated, low cost, and the interpretation of test results remains at the manual stage, that is, the test results of the reagent card are interpreted by human eyes, with a low degree of automation, and it is very likely that human misjudgment of the test results will occur. In addition, the test results cannot be traced. The microcolumn gel method is currently the blood typing technology recommended by the World Health Organization. The microcolumn gel reagent card consists of a microcolumn tube and a nameplate: the microcolumn tube is the reaction container for blood typing. During the test, red blood cells or serum are added to the upper part of the microcolumn tube. The test result is determined by analyzing the distribution of red blood cells in the microcolumn tube after the reaction; the nameplate records the type and information of the blood typing reagent card. The microplate method has the characteristics of high throughput and automation, and is widely used in blood stations and large hospitals, but the raw materials of microcolumn gel particles are mostly imported and are currently monopolized by foreign countries.

[0003] In recent years, routine blood typing tests for blood donors at all levels of blood centers and blood stations in my country have used both liquid-paper disc reagents and microplate methods. In blood transfusion departments and blood banks at all levels of hospitals, routine blood typing tests for patients and blood donors (waiting for blood transfusion) are mostly carried out using two types of detection technologies, namely microcolumn gel immunoassay (MGIA) and liquid-paper disc reagents. A few large hospitals use three technologies, namely liquid-paper disc reagents, microplate methods and MGIA. The advantages and disadvantages of these methods are shown in Table 1 below.

[0004] Table 1. Comparison of the advantages and disadvantages of several blood typing methods

[0005]

[0006] Therefore, there is an urgent need for a new red blood cell typing test solution that is easy to operate, accurate, highly sensitive, and has independent and controllable raw materials. Summary of the invention

[0007] The technical problem to be solved by the present invention is to provide a red blood cell type detection and antibody screening and identification system and method in view of the deficiencies in the above-mentioned prior art.

[0008] In order to solve the above technical problems, the technical solution adopted by the present invention is: a red blood cell blood typing detection and antibody screening identification system, comprising: a positive typing detection liquid phase exclusion chromatography separation medium, a reverse typing detection liquid phase exclusion chromatography separation medium, an antibody screening liquid phase exclusion chromatography separation medium and a microcolumn card;

[0009] The positive typing detection liquid phase exclusion chromatography separation medium, the reverse typing detection liquid phase exclusion chromatography separation medium, and the antibody screening liquid phase exclusion chromatography separation medium are respectively filled in microcolumn cards, and the observation window of the microcolumn card is observed after centrifugation to independently realize the positive typing detection of red blood cell blood type, the reverse typing detection of red blood cell blood type and the screening and identification of red blood cell antibodies.

[0010] Preferably, the microcolumn card is provided with a plurality of mutually independent microcolumns, the microcolumn comprises a reaction chamber, a separation chamber and an observation window arranged in sequence from top to bottom, and the reaction chamber is provided with a sealing cover;

[0011] The width of the reaction chamber is greater than that of the separation chamber, and the reaction chamber and the separation chamber are transitionally connected via a tapered portion.

[0012] Preferably, the positive typing detection liquid exclusion chromatography separation medium is prepared by the following method:

[0013] Sucrose, polysucrose, diatrizoate, PEG, and bovine serum albumin are added to 0.01M phosphate buffer, and then anti-A, anti-B or anti-D is added, and the mixture is evenly mixed to obtain a positive typing detection liquid exclusion chromatography separation medium.

[0014] Preferably, the mass ratio of each component in the liquid exclusion chromatography separation medium for positive typing detection is as follows: 30% sucrose, 2% polysucrose, 1% diatrizoate, 1% PEG, and 2% bovine serum albumin;

[0015] The positive typing test for anti-A, anti-B or anti-D antibodies in the liquid exclusion chromatography separation medium was 1:128.

[0016] The specific gravity of the liquid exclusion chromatography separation medium for positive characterization was 1.10 and the pH was 7.0.

[0017] Preferably, the reverse typing detection liquid exclusion chromatography separation medium is prepared by the following method:

[0018] Sucrose, polysucrose, diatrizoate, PEG, and bovine serum albumin are added into 0.01 M phosphate buffer, and mixed evenly to obtain a positive typing detection liquid exclusion chromatography separation medium.

[0019] Preferably, the mass ratio of the components in the liquid exclusion chromatography separation medium for reverse typing detection is as follows: 30% sucrose, 2% polysucrose, 1% diatrizoate, 1% PEG, and 2% bovine serum albumin;

[0020] The specific gravity of the liquid exclusion chromatography separation medium for reverse typing detection was 1.10 and the pH was 7.0.

[0021] Preferably, the antibody screening liquid exclusion chromatography separation medium is prepared by the following method:

[0022] Sucrose, polysucrose, diatrizoate, PEG, and bovine serum albumin are added to 0.01 M phosphate buffer, and then goat anti-human IgG is added, and after mixing evenly, a positive typing detection liquid exclusion chromatography separation medium is obtained;

[0023] The mass ratio of each component in the antibody screening liquid exclusion chromatography separation medium is as follows: 30% sucrose, 2% polysucrose, 1% diatrizoate, 1% PEG, and 2% bovine serum albumin;

[0024] In the antibody screening liquid exclusion chromatography separation medium, the titer of goat anti-human IgG was 1:512;

[0025] The antibody screening liquid exclusion chromatography separation medium has a specific gravity of 1.10 and a pH of 7.0.

[0026] Preferably, the method for performing positive typing of red blood cells in the system is:

[0027] S1-1, filling the positive typing test liquid exclusion chromatography separation medium into the microcolumn of the microcolumn card, with the filling volume in a single microcolumn being 70-280 μL;

[0028] S1-2, centrifuge at 1500-6000 rpm for 15-60 s;

[0029] S1-3, dilute the red blood cells to be tested with physiological saline to a mass concentration of 0.25%-1% and then add them to the microcolumn. The amount added to a single microcolumn is 25-100 μL;

[0030] S1-4, centrifuge at 450-1800 rpm for 1-4 min, then centrifuge at 900-3600 rpm for 1.5-6 min, and observe the results through the observation window: if there are agglutinated red blood cells in the observation window, it is a positive result, otherwise it is a negative result.

[0031] Preferably, the method for performing red blood cell reverse typing detection in the system is:

[0032] S2-1, filling the reverse typing detection liquid exclusion chromatography separation medium into the microcolumn of the microcolumn card, with the filling volume in a single microcolumn being 70-280 μL;

[0033] S2-2, centrifuge at 1500-6000 rpm for 15-60 s;

[0034] S2-3, prepare a reverse red blood cell typing reagent with a mass concentration of 0.25%-1% using normal saline;

[0035] S2-4, add the plasma or serum to be tested into the microcolumn, the amount added to a single microcolumn is 25-100 μL;

[0036] S2-5, add 25-100 μL of the anti-erythrocyte typing reagent prepared in step S2-3 into the microcolumn;

[0037] S2-6, centrifuge at 450-1800 rpm for 1-4 min, then centrifuge at 900-3600 rpm for 1.5-6 min, and observe the results through the observation window: if there are agglutinated red blood cells in the observation window, it is a positive result, otherwise it is a negative result.

[0038] Preferably, the method for screening and identifying red blood cell antibodies in the system is:

[0039] S3-1, filling the antibody screening liquid exclusion chromatography separation medium into the microcolumn of the microcolumn card, with the filling volume in a single microcolumn being 70-280 μL;

[0040] S3-2, centrifuge at 1500-6000 rpm for 15-60 s;

[0041] S3-3, dilute the screening cells or spectrum cells with physiological saline to a mass concentration of 0.25%-1%;

[0042] S3-4, add the plasma or serum to be tested into the microcolumn, the amount added to a single microcolumn is 25-100 μL;

[0043] S3-5, adding 25-100 μL of the screening cells or spectrum cells prepared in step S3-3 into the microcolumn;

[0044] S3-6, centrifuge at 450-1800 rpm for 1-4 min, then centrifuge at 900-3600 rpm for 1.5-6 min, and observe the results through the observation window: if there are agglutinated red blood cells in the observation window, it is a positive result, otherwise it is a negative result.

[0045] The principle of the present invention is the liquid exclusion chromatography principle, that is, in the liquid exclusion chromatography separation medium, the red blood cell antigen reacts with the corresponding antibody to form an agglutinated red blood cell immune complex. Under the action of a certain centrifugal force, the complex passes through the liquid exclusion chromatography separation medium and is deposited at the bottom of the microcolumn card observation window or suspended in the observation window glue, which is a positive result; if there is no corresponding antibody binding, the red blood cell immune complex cannot be formed. Under the same centrifugal force, the dispersed red blood cells are suspended in the upper layer of the liquid exclusion chromatography separation medium in the separation chamber, and there is no cell deposition in the observation window, which is a negative result.

[0046] The beneficial effects of the present invention are:

[0047] The present invention provides a red blood cell blood type detection and antibody screening and identification system and method, which can realize red blood cell positive typing, reverse typing, antibody screening, antibody identification, etc. The detection scheme of the present invention is conducive to promoting clinical red blood cell blood type compatibility detection, and can realize rapid and accurate detection of red blood cell ABO, Rh, MNS, Kell, Kidd and other blood types of various blood type systems, and can also select different detection items according to POCT detection (point-of-care detection) needs, so as to be suitable for various blood type detection situations such as hospitals, families and field emergencies.

[0048] The detection process of the present invention can achieve red blood cell typing and cross-matching without washing. At the same time, the vertical liquid phase detection system uses a liquid exclusion chromatography separation medium to separate and avoid false positive reactions caused by fibrinogen. The operation is simple and the results are intuitive and reliable. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 It is a schematic diagram of the structure of the microcolumn card of the present invention;

[0050] Figure 2 It is a display diagram of the detection results in Example 2 of the present invention;

[0051] Figure 3 It is a display diagram of the detection results in Example 3 of the present invention;

[0052] Figure 4 This is a graph showing the detection results in Example 4 of the present invention.

[0053] Description of reference numerals:

[0054] 1—microcolumn card; 10—microcolumn; 11—reaction chamber; 12—separation chamber; 13—observation window; 14—sealing cover. DETAILED DESCRIPTION

[0055] The present invention is further described in detail below in conjunction with embodiments so that those skilled in the art can implement the invention with reference to the description.

[0056] It should be understood that the terms such as “having”, “including” and “comprising” used herein do not exclude the existence or addition of one or more other elements or combinations thereof.

[0057] The test methods used in the following examples are conventional methods unless otherwise specified. The materials and reagents used in the following examples are all commercially available unless otherwise specified. In the following examples, if no specific conditions are specified, the experiments were carried out under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used, if the manufacturer is not specified, are all conventional products that can be purchased commercially.

[0058] Example 1

[0059] A red blood cell blood typing detection and antibody screening and identification system, comprising: a positive typing detection liquid phase exclusion chromatography separation medium, a reverse typing detection liquid phase exclusion chromatography separation medium, an antibody screening liquid phase exclusion chromatography separation medium and a microcolumn card 1;

[0060] The positive typing detection liquid phase exclusion chromatography separation medium, the reverse typing detection liquid phase exclusion chromatography separation medium, and the antibody screening liquid phase exclusion chromatography separation medium are respectively filled in the microcolumn card 1, and the observation window 13 of the microcolumn card 1 is observed after centrifugation to independently realize the positive typing detection of red blood cell blood type, the reverse typing detection of red blood cell blood type and the screening and identification of red blood cell antibodies.

[0061] In this embodiment, a plurality of mutually independent microcolumns 10 are arranged on the microcolumn card 1. The microcolumn 10 includes a reaction chamber 11, a separation chamber 12 and an observation window 13 arranged in sequence from top to bottom. A sealing cover 14 is arranged on the reaction chamber 11. The width of the reaction chamber 11 is greater than that of the separation chamber 12. The reaction chamber 11 and the separation chamber 12 are transitionally connected by a tapered portion. More specifically, the microcolumn card 1 used in this embodiment has the same structure and principle as the zone distribution detection card in the platelet anti-human globulin experimental immune zone distribution detection method of patent CN112379100B.

[0062] In this embodiment, the positive type detection liquid phase exclusion chromatography separation medium is prepared by the following method:

[0063] Sucrose, polysucrose, diatrizoate, PEG, and bovine serum albumin are added to 0.01M phosphate buffer, and then anti-A, anti-B or anti-D is added, and the mixture is evenly mixed to obtain a positive typing detection liquid exclusion chromatography separation medium.

[0064] The mass ratio of each component in the liquid exclusion chromatography separation medium for positive typing test is as follows: 30% sucrose, 2% polysucrose, 1% diatrizoate, 1% PEG (molecular weight 4000), and 2% bovine serum albumin;

[0065] The positive typing test for anti-A, anti-B or anti-D antibodies in the liquid exclusion chromatography separation medium was 1:128.

[0066] The specific gravity of the liquid exclusion chromatography separation medium for positive characterization was 1.10 and the pH was 7.0.

[0067] In this embodiment, the reverse typing detection liquid exclusion chromatography separation medium is prepared by the following method:

[0068] Sucrose, polysucrose, diatrizoate, PEG, and bovine serum albumin are added into 0.01 M phosphate buffer, and mixed evenly to obtain a positive typing detection liquid exclusion chromatography separation medium.

[0069] The mass ratio of each component in the liquid exclusion chromatography separation medium for reverse typing detection is as follows: 30% sucrose, 2% polysucrose, 1% diatrizoate, 1% PEG (molecular weight 4000), and 2% bovine serum albumin;

[0070] The specific gravity of the liquid exclusion chromatography separation medium for reverse typing detection was 1.10 and the pH was 7.0.

[0071] In this embodiment, the antibody screening liquid exclusion chromatography separation medium is prepared by the following method:

[0072] Sucrose, polysucrose, diatrizoate, PEG, and bovine serum albumin are added to 0.01 M phosphate buffer, and then goat anti-human IgG is added, and after mixing evenly, a positive typing detection liquid exclusion chromatography separation medium is obtained;

[0073] The mass ratio of each component in the antibody screening liquid exclusion chromatography separation medium is as follows: 30% sucrose, 2% polysucrose, 1% diatrizoate, 1% PEG (molecular weight 4000), and 2% bovine serum albumin;

[0074] In the antibody screening liquid exclusion chromatography separation medium, the titer of goat anti-human IgG was 1:512;

[0075] The antibody screening liquid exclusion chromatography separation medium has a specific gravity of 1.10 and a pH of 7.0.

[0076] Example 2

[0077] The red blood cell blood type detection and antibody screening and identification system of Example 1 is used to perform red blood cell blood type positive typing detection, and the specific method is as follows:

[0078] S1-1, filling the positive typing detection liquid exclusion chromatography separation medium into the microcolumn 10 of the microcolumn card 1, with the filling volume in a single microcolumn 10 being 140 μL;

[0079] S1-2, centrifuge at 3000 rpm for 30 s;

[0080] S1-3, dilute the red blood cells to be tested with physiological saline (0.9wt% sodium chloride) to a mass concentration of 0.5% and then add them to the microcolumn 10, with the amount added to a single microcolumn 10 being 50 μL;

[0081] S1-4, centrifuge at 900rpm for 2min, then centrifuge at 1800rpm for 3min, and observe the result through the observation window 13: if there are agglutinated red blood cells in the observation window 13, it is a positive result, otherwise it is a negative result. Agglutinated red blood cells that have undergone antigen-antibody reaction pass through the medium under the action of centrifugal force, and are distributed in the observation window 13 of the microcolumn card 1 or deposited on the bottom edge. Agglutinated red blood cells can be observed in the observation window 13, which is a positive result; on the contrary, red blood cells that have not undergone antigen-antibody reaction cannot pass through the medium under the action of centrifugal force and are distributed in the separation chamber 12 medium of the microcolumn card 1, which is a negative result. Figure 2 , are the test results. It can be seen that the red blood cells of the first blood sample reacted positively with anti-A and anti-D, indicating that its positive typing result was type A RhD(+), the red blood cells of the second blood sample reacted positively with anti-B and anti-D, indicating that its positive typing result was type B RhD(+), and the red blood cells of the third blood sample only reacted positively with anti-D, indicating that its positive typing result was type O RhD(+).

[0082] Example 3

[0083] The red blood cell blood type detection and antibody screening and identification system of Example 1 is used to perform red blood cell blood type reverse typing detection, and the specific method is as follows:

[0084] S2-1, filling the reverse typing detection liquid exclusion chromatography separation medium into the microcolumn 10 of the microcolumn card 1, with the filling volume of a single microcolumn 10 being 140 μL;

[0085] S2-2, centrifuge at 3000 rpm for 30 s;

[0086] S2-3, prepare a 0.5% reverse red blood cell typing reagent (commonly available commercially available human ABO blood type reverse red blood cell typing reagent) with physiological saline;

[0087] S2-4, adding the plasma or serum to be tested into the microcolumn 10, with the amount added to a single microcolumn 10 being 50 μL;

[0088] S2-5, adding 50 μL of the anti-erythrocyte typing reagent prepared in step S2-3 into the microcolumn 10;

[0089] S2-6, centrifuge at 900 rpm for 2 min, then centrifuge at 1800 rpm for 3 min, and observe the result through the observation window 13: if there are agglutinated red blood cells in the observation window 13, it is a positive result, otherwise it is a negative result.

[0090] Agglutinated red blood cells that have undergone antigen-antibody reaction pass through the medium under the action of centrifugal force and are distributed in the observation window 13 of the microcolumn card 1 or deposited on the lower edge. Agglutinated red blood cells can be observed in the observation window 13, which is a positive result. On the contrary, red blood cells that have not undergone antigen-antibody reaction cannot pass through the medium under the action of centrifugal force and are distributed in the upper layer of the medium in the separation chamber 12 of the microcolumn card 1, which is a negative result. Figure 3 , is the test result, it can be seen

[0091] The first plasma reacts positively with the anti-typing reagent type B red blood cells, indicating that it is blood type A, and the second plasma reacts positively with the anti-typing reagent type A red blood cells, indicating that it is blood type B. The second plasma reacts positively with both the anti-typing reagent type A and type B red blood cells, indicating that it is blood type O.

[0092] Example 4

[0093] The red blood cell blood type detection and antibody screening and identification system of Example 1 is used to perform red blood cell antibody screening and identification, and the specific method is as follows:

[0094] S3-1, filling the antibody screening liquid exclusion chromatography separation medium into the microcolumn 10 of the microcolumn card 1, with the filling volume of a single microcolumn 10 being 140 μL;

[0095] S3-2, centrifuge at 3000 rpm for 30 s;

[0096] S3-3, dilute the screening cells or spectrum cells with physiological saline to a mass concentration of 0.5%;

[0097] S3-4, adding the plasma or serum to be tested into the microcolumn 10, with the amount added to a single microcolumn 10 being 50 μL;

[0098] S3-5, adding 50 μL of the screening cells or spectrum cells (irregular antibody screening red blood cell reagent, antibody identification spectrum cell reagent) prepared in step S1-3 to the microcolumn 10;

[0099] S3-6, centrifuge at 900rpm for 2min, then centrifuge at 1800rpm for 3min, and observe the result through the observation window 13: if there are agglutinated red blood cells in the observation window 13, it is a positive result, otherwise it is a negative result. Agglutinated red blood cells that have undergone antigen-antibody reaction pass through the medium under the action of centrifugal force, and are distributed in the observation window 13 of the microcolumn card 1 or deposited on the bottom edge. Agglutinated red blood cells can be observed in the observation window 13, which is a positive result; on the contrary, red blood cells that have not undergone antigen-antibody reaction cannot pass through the medium under the action of centrifugal force and are distributed in the separation chamber 12 medium of the microcolumn card 1, which is a negative result. Figure 4 , is the test result, it can be seen that:

[0100] When the red blood cell complex forms agglutination and completely settles at the bottom edge of the microcolumn card observation window, the result is a strong positive reaction, expressed as "4+"; when most of the red blood cell complexes form agglutination and settle at the bottom edge of the microcolumn card observation window, very few agglutinations are dispersed in the medium of the observation window, the result is a relatively strong positive reaction, expressed as "3+"; when a small number of red blood cell complexes form agglutination and deposit at the bottom edge of the observation window, part of the agglutination is dispersed in the medium of the observation window, and some non-agglutinated red blood cells are located in the separation chamber, the result is a weak positive reaction, expressed as "2+"; when very few red blood cell complexes form agglutination and deposit at the bottom edge of the observation window, a small part of the agglutination is dispersed in the medium of the observation window, and most of the non-agglutinated red blood cells are located in the separation chamber, the result is a weak positive reaction, expressed as "1+"; when only a very small number of red blood cells are located at the bottom of the separation chamber of the microcolumn card and the top of the observation window, it is impossible to tell whether the red blood cells are agglutinated by the naked eye, and the result is suspicious, expressed as " + "; At this time, other methods are needed to re-test and confirm. When there are no red blood cells in the observation window of the micro-column card, the unagglutinated red blood cells are all distributed in the separation chamber of the micro-column card, and the result is a negative reaction, which is expressed as "-".

[0101] Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and the implementation modes. They can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to specific details.

Claims

1. A red blood cell type detection and antibody screening and identification system, characterized in that: include: Positive typing detection liquid exclusion chromatography separation medium, reverse typing detection liquid exclusion chromatography separation medium, antibody screening liquid exclusion chromatography separation medium and microcolumn card; The positive typing detection liquid phase exclusion chromatography separation medium, the reverse typing detection liquid phase exclusion chromatography separation medium, and the antibody screening liquid phase exclusion chromatography separation medium are respectively filled in microcolumn cards, and the observation window of the microcolumn card is observed after centrifugation to independently realize the positive typing detection of red blood cell blood type, the reverse typing detection of red blood cell blood type and the screening and identification of red blood cell antibodies.

2. The red blood cell type detection and antibody screening and identification system according to claim 1, characterized in that: The microcolumn card is provided with a plurality of mutually independent microcolumns, the microcolumns comprising a reaction chamber, a separation chamber and an observation window arranged in sequence from top to bottom, and a sealing cover is provided on the reaction chamber; The width of the reaction chamber is greater than that of the separation chamber, and the reaction chamber and the separation chamber are transitionally connected via a tapered portion.

3. The red blood cell type detection and antibody screening and identification system according to claim 1, characterized in that: The positive typing detection liquid exclusion chromatography separation medium is prepared by the following method: Sucrose, polysucrose, diatrizoate, PEG, and bovine serum albumin are added to 0.01M phosphate buffer, and then anti-A, anti-B or anti-D is added, and the mixture is evenly mixed to obtain a positive typing detection liquid exclusion chromatography separation medium.

4. The red blood cell type detection and antibody screening and identification system according to claim 3, characterized in that: The mass ratio of each component in the liquid exclusion chromatography separation medium for positive typing test is as follows: 30% sucrose, 2% polysucrose, 1% diatrizoate, 1% PEG, and 2% bovine serum albumin; The positive typing test for the titer of anti-A, anti-B or anti-D antibodies in the liquid exclusion chromatography separation medium was 1:

128. The specific gravity of the liquid exclusion chromatography separation medium for positive characterization was 1.10 and the pH was 7.

0.

5. The red blood cell type detection and antibody screening and identification system according to claim 1, characterized in that: The reverse typing detection liquid exclusion chromatography separation medium was prepared by the following method: Sucrose, polysucrose, diatrizoate, PEG, and bovine serum albumin are added into 0.01 M phosphate buffer, and mixed evenly to obtain a positive typing detection liquid exclusion chromatography separation medium.

6. The red blood cell type detection and antibody screening and identification system according to claim 5, characterized in that: The mass ratio of each component in the liquid exclusion chromatography separation medium for reverse typing detection is as follows: 30% sucrose, 2% polysucrose, 1% diatrizoate, 1% PEG, and 2% bovine serum albumin; The specific gravity of the liquid exclusion chromatography separation medium for reverse typing detection was 1.10 and the pH was 7.

0.

7. The red blood cell type detection and antibody screening and identification system according to claim 1, characterized in that: The antibody screening liquid phase separation medium was prepared by the following method: Sucrose, polysucrose, diatrizoate, PEG, and bovine serum albumin are added to 0.01M phosphate buffer, and then goat anti-human IgG is added, and after mixing evenly, an antibody screening liquid exclusion chromatography separation medium is obtained; The mass ratio of each component in the antibody screening liquid exclusion chromatography separation medium is as follows: 30% sucrose, 2% polysucrose, 1% diatrizoate, 1% PEG, and 2% bovine serum albumin; In the antibody screening liquid exclusion chromatography separation medium, the titer of goat anti-human IgG was 1:512; The specific gravity of the antibody screening liquid exclusion chromatography separation medium is 1.10 and the pH is 7.

0.

8. The red blood cell type detection and antibody screening and identification system according to claim 4, characterized in that: The method for the system to perform positive typing of red blood cells is: S1-1, filling the positive typing test liquid exclusion chromatography separation medium into the microcolumn of the microcolumn card, with the filling volume in a single microcolumn being 70-280 μL; S1-2, centrifuge at 1500-6000 rpm for 15-60 s; S1-3, dilute the red blood cells to be tested with physiological saline to a mass concentration of 0.25%-1% and then add them to the microcolumn. The amount added to a single microcolumn is 25-100 μL; S1-4, centrifuge at 450-1800 rpm for 1-4 min, then centrifuge at 900-3600 rpm for 1.5-6 min, and observe the results through the observation window: if there are agglutinated red blood cells in the observation window, it is a positive result, otherwise it is a negative result.

9. The red blood cell type detection and antibody screening and identification system according to claim 6, characterized in that: The method for the system to perform red blood cell reverse typing detection is: S2-1, filling the reverse typing detection liquid exclusion chromatography separation medium into the microcolumn of the microcolumn card, with the filling volume in a single microcolumn being 70-280 μL; S2-2, centrifuge at 1500-6000 rpm for 15-60 s; S2-3, prepare a reverse red blood cell typing reagent with a mass concentration of 0.25%-1% using normal saline; S2-4, add the plasma or serum to be tested into the microcolumn, the amount added to a single microcolumn is 25-100 μL; S2-5, add 25-100 μL of the anti-erythrocyte typing reagent prepared in step S2-3 into the microcolumn; S2-6, centrifuge at 450-1800 rpm for 1-4 min, then centrifuge at 900-3600 rpm for 1.5-6 min, and observe the results through the observation window: if there are agglutinated red blood cells in the observation window, it is a positive result, otherwise it is a negative result.

10. The red blood cell type detection and antibody screening and identification system according to claim 7, characterized in that: The method for the system to screen and identify red blood cell antibodies is: S3-1, filling the antibody screening liquid exclusion chromatography separation medium into the microcolumn of the microcolumn card, with the filling volume in a single microcolumn being 70-280 μL; S3-2, centrifuge at 1500-6000 rpm for 15-60 s; S3-3, dilute the screening cells or spectrum cells with physiological saline to a mass concentration of 0.25%-1%; S3-4, add the plasma or serum to be tested into the microcolumn, the amount added to a single microcolumn is 25-100 μL; S3-5, adding 25-100 μL of the screening cells or spectrum cells prepared in step S3-3 into the microcolumn; S3-6, centrifuge at 450-1800 rpm for 1-4 min, then centrifuge at 900-3600 rpm for 1.5-6 min, and observe the results through the observation window: if there are agglutinated red blood cells in the observation window, it is a positive result, otherwise it is a negative result.

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