Application of non-ionic sugar aqueous solution in blood group irregular antibody detection and detection method

Abstract

The invention provides application of a non-ionic sugar aqueous solution in blood group irregular antibody detection and a detection method, and relates to the field of medical examination. The ion-free sugar aqueous solution is a 10% sucrose deionized aqueous solution or a trehalose aqueous solution, and the method for detecting the irregular antibody in the blood comprises the following steps: adding the ion-free sugar aqueous solution into a red blood cell suspension and to-be-detected plasma mixture, centrifuging, removing supernate, adding normal saline, and observing whether non-specific agglutinates are dissipated within 1 minute. The ion-free sugar aqueous solution is used as a reinforcing agent, the components are simple, the preparation is easy, the detection capability and the detection performance on irregular antibodies in blood are strong, and the false positive and false negative probability is low.

Description

Technical Field

[0001] The invention relates to the field of medical testing, and in particular to an application of a non-ionized sugar aqueous solution in the detection of irregular blood type antibodies and a detection method. Background Art

[0002] Incomplete antibodies refer to antibodies that can bind to corresponding antigens in saline media but cannot produce visible agglutination reactions. Incomplete antibodies are mainly IgG antibodies. After binding to the corresponding antigens on red blood cells, special methods, such as anti-human globulin method, polybrene method, and enzyme method, must be used to cause sensitized red blood cells to undergo agglutination reactions, thereby detecting such antibodies. Irregular antibodies refer to blood type antibodies that do not conform to the "Landsteiner rule". Complete antibodies and incomplete antibodies may exist alone or simultaneously. They are currently the main factor leading to allogeneic blood transfusion incompatibility reactions and neonatal hemolytic disease in clinical practice. Therefore, in pre-transfusion compatibility testing and neonatal hemolytic disease-related testing, we need to use various test methods to screen and identify irregular antibodies in ex vivo blood samples.

[0003] However, the results of irregular antibody screening and specific identification are often restricted by various practical factors, such as the sensitivity of the detection method, the difference in the optimal reaction conditions of different antibodies, and the "dose effect" caused by heterozygous reagent cells, etc. False negative results cause test operators to mistakenly send incompatible blood to clinical patients for transfusion, resulting in ineffective transfusion or adverse reactions; false positive results also lead to the waste of test reagents, consumables, human resources and time, delaying the use of blood for clinical patients.

[0004] According to the national clinical laboratory operation procedures and the relevant chapters in the AABB manual, combined with the actual clinical application, it can be concluded that the methods used in the cross-matching test, irregular antibody screening and identification test that are currently widely carried out are mainly: saline test tube method (NS), polybrene reagent test tube method (MPT), anti-human globulin reagent test tube method (IAT), anti-human globulin reagent microcolumn gel card method (MGT), etc. Among them, NS is mainly used to detect IgM complete antibodies, and MPT, IAT and MGT are mainly used to detect IgG incomplete antibodies. The above detection methods have their own advantages and disadvantages. No method can guarantee that irregular antibodies will not be missed. In addition, the clinical blood transfusion technical specifications also require that at least two methods should be used for mutual verification to prevent the missed detection of low-titer irregular antibodies. The AABB manual introduces many reagents and methods for enhancing red blood cell antigen and antibody reactions. However, many of these reagents and methods have not yet been widely used in clinical practice, which may cause certain hidden dangers to the safety of blood transfusion in clinical patients. Summary of the invention

[0005] The purpose of the present invention is to provide an application of an ion-free sugar aqueous solution in the detection of irregular blood type antibodies. The ion-free sugar aqueous solution is used as an enhancer, has simple ingredients, is easy to configure, has strong detection ability and test performance for irregular antibodies in the blood, and has a low probability of false positives and false negatives.

[0006] Another object of the present invention is to provide a method for detecting irregular antibodies, which adopts the traditional test tube method, uses an ion-free sugar aqueous solution as an enhancer to promote non-specific agglutination of red blood cells, and then uses physiological saline as a deaggregating agent to dissipate the non-specific agglutination of red blood cells. The detection method has high accuracy and is easy to operate.

[0007] The third aspect of the present invention provides the application of the ion-free sugar aqueous solution as an enhancer in the irregular antibody detection of the microcolumn gel method. The traditional test tube method is first adopted, and the ion-free sugar aqueous solution is used as an enhancer to promote the non-specific agglutination of red blood cells. Then, physiological saline is used as a disaggregating agent to dissipate the non-specific agglutination of red blood cells. Thereafter, the red blood cell suspension is added to the microcolumn gel anti-human globulin card and directly centrifuged. This detection method can omit the step of incubating at 37°C in the traditional MGT method, thereby saving operation time and having a high accuracy rate.

[0008] The present invention solves the technical problem by adopting the following technical solutions.

[0009] On the one hand, an embodiment of the present invention provides an application of an ion-free sugar aqueous solution in the detection of irregular blood type antibodies.

[0010] In some embodiments of the present invention, the ion-free sugar aqueous solution is used in the detection of microcolumn gel anti-human globulin.

[0011] In some embodiments of the present invention, the ion-free sugar aqueous solution is a sucrose aqueous solution, a trehalose aqueous solution or a mixture of the two.

[0012] In some embodiments of the present invention, the mass fraction of the ion-free sugar aqueous solution is 10%.

[0013] In some embodiments of the present invention, the sucrose solid used in the preparation of the sucrose aqueous solution is of analytically pure AR grade; the trehalose solid used in the preparation of the trehalose aqueous solution is of analytically pure AR grade.

[0014] In some embodiments of the present invention, the conductivity (25° C.) of deionized water used in the preparation of the sucrose aqueous solution and the trehalose aqueous solution is ≤0.055 μs / cm.

[0015] In some embodiments of the present invention, the method for detecting irregular blood type antibodies using the ion-free sugar aqueous solution in a traditional test tube method comprises the following steps: adding the ion-free sugar aqueous solution to a mixture of a red blood cell suspension expressing a specific antigen and the plasma to be tested, centrifuging, removing the supernatant, and then adding physiological saline to observe whether the nonspecific agglutinates dissipate within 1 minute.

[0016] In some embodiments of the present invention, the irregular antibody is an Rh blood type anti-D or anti-E incomplete antibody.

[0017] In some embodiments of the present invention, the centrifugal force during centrifugation is 1000 g, and the centrifugation time is 1-2 min. Preferably, the centrifugation time is 1 min.

[0018] On the other hand, the embodiment of the present invention provides a method for using an ion-free sugar aqueous solution as a reaction enhancer in the microcolumn gel anti-human globulin method to detect irregular antibodies in blood, comprising the following steps: adding an ion-free sugar aqueous solution to a mixture of a red blood cell suspension expressing a specific antigen and a plasma to be tested, centrifuging, removing the supernatant, and then adding physiological saline to observe whether the nonspecific agglutinate dissipates within 1 minute. Thereafter, resuspending and adding to a microcolumn gel anti-human globulin card and directly centrifuging to observe the red blood cell agglutination degree in the microcolumn.

[0019] Compared with the prior art, the embodiments of the present invention have at least the following advantages or beneficial effects:

[0020] The application of the non-ionized sugar aqueous solution provided by the present invention in the detection of irregular antibodies by the traditional test tube method, the non-ionized sugar aqueous solution has a simple composition formula and easy test steps, and can replace the polybrene method for the detection of incomplete antibodies of Rh blood type, and is suitable for promotion and application in primary clinical laboratories. Specifically, the non-ionized sugar aqueous solution is used as an enhancer to promote the non-specific binding and aggregation of red blood cells. The ions cannot be selectively aggregated at the solid-liquid interface in the sucrose deionized water solution system, thereby preventing the red blood cells from being charged on the surface and hindering the formation of the red blood cells' zeta potential, and promoting the red blood cells antigen to bind to its specific IgG antibody. It has been confirmed by experiments that this method can significantly reduce the agglutination caused by IgM antibodies and enhance the detection of IgG antibodies.

[0021] The incomplete antibody detection method provided by the present invention is compared with the traditional polybrene reagent test tube method. The first step of the polybrene reagent test tube method is to mix the plasma to be tested with the red blood cells expressing the known antigen, and then add the low ion medium (LIM) and the polybrene reagent, and then centrifuge to determine whether the red blood cells are agglutinated to determine whether the incomplete antibodies exist. Therefore, whether the red blood cells obtained after centrifugation are agglutinated cannot be determined due to the action of polybrene or due to the interference of IgM blood type antibodies in the plasma to be tested. It is necessary to use the test results of the saline test tube method as a reference, which adds additional inspection operations. In addition, since a strong cationic polymer polybrene is used to promote non-specific agglutination of red blood cells, a specific citrate solution is required in the second step for depolymerization, and the composition of citrate is complex and not easy to configure, and the pH value of the antigen-antibody reaction environment is changed, which may cause some antibodies to be missed. The incomplete antibody detection method provided by the present invention verifies whether the red blood cells produce specific agglutination under the action of the non-ionized sugar aqueous solution in the subsequent step, without being interfered by the IgM complete antibody, and only needs to use NaCl in the second step to provide free electrons in the solution, restore the zeta potential of the red blood cells, and achieve deaggregation, without changing the pH value of the antigen-antibody reaction environment. The reagent components used are simple and easy to configure.

[0022] In addition, in the polycondensation test to microcolumn gel anti-human globulin test, since the LIM solution used in the first step of the polycondensation test can interfere with the anti-human globulin reaction, if the red blood cell plasma mixture after the second step of depolymerization is directly added to the microcolumn gel anti-human globulin card for reaction, a false positive result may be caused, but the present invention does not have this problem. At the same time, the use of an ion-free sugar aqueous solution as an enhancer can omit the step of incubating the antigen and antibody at 37°C in the microcolumn gel anti-human globulin test, saving operation time and improving the detection rate of blood type antibodies. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0024] Figure 1 is the IgG anti-D antibody titer in the embodiment of the present invention;

[0025] Figure 2 is the IgG anti-E antibody titer in the embodiment of the present invention;

[0026] Figure 3The comparison of the detection effect between the sucrose aqueous solution method and the polybrene method in the embodiment of the present invention;

[0027] Figure 4 The results of the microcolumn gel anti-human globulin test for clinical patients are as follows;

[0028] Figure 5 The results of the microcolumn gel anti-human globulin test for clinical patients are as follows;

[0029] Figure 6 The results of microcolumn gel anti-human globulin test for clinical patients are three. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0031] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other. The present invention will be described in detail below with reference to specific embodiments.

[0032] The embodiment of the present invention provides an application of an ion-free sugar aqueous solution in the detection of irregular blood type antibodies.

[0033] More preferably, the ion-free sugar aqueous solution is used in the detection of irregular blood type antibodies by the traditional test tube method.

[0034] More preferably, the ion-free sugar aqueous solution is used in the detection of microcolumn gel anti-human globulin.

[0035] The main components of low ion medium (LIM) are glucose, Na 2 EDTA·2H 2 O, NaOH, low ionic strength solution (LISS) mainly contains glycine, Na 2 HPO 4 , KH 2 PO 4 , NaCl. LIM is suitable for the first step of the polybrene test to provide a medium environment for specific proteins and free proteins to approach red blood cells; while LISS is suitable for the enhancement of the anti-human globulin test, and its function is to reduce the ions in the medium, reduce the zeta potential of red blood cells, and promote the binding of red blood cell antigens to their specific antibodies. The two have different components and scopes of application and cannot be replaced by each other. For example, LIM used in the anti-human globulin test tube method and the cassette method can lead to false positives and interfere with the result determination; while LISS used in the low-ion polybrene test can easily lead to false negatives.

[0036] The sucrose aqueous solution provided in the embodiment of the present invention comprises sucrose and deionized water, wherein one molecule of sucrose is formed by one molecule of glucose and one molecule of fructose connected by chemical bonds, and has excellent solubility. In in vitro tests, it can be re-dissociated into glucose and fructose under the action of acid or enzyme. Therefore, theoretically, the sucrose molecules dissolved in deionized water form a non-ionic sucrose solution (NISS). The solid-liquid interface in the NISS system cannot selectively aggregate ions, thereby preventing the red blood cell surface from being charged, hindering the formation of the red blood cell zeta potential, and promoting the red blood cell antigen to bind to its specific IgG antibody. Therefore, in principle, it is closer to LISS (the main components are glycine, Na 2 HPO 4 , KH 2 PO 4 , NaCl) mechanism of action. 2 EDTA·2H 2 O is one of the key components of low-ion media. The ion-free sugar aqueous solution provided by the present invention is only suitable for plasma samples of whole blood anticoagulated with K2EDTA. The EDTA component in the reaction system depends on the anticoagulant dissolved in the blood collection tube in the plasma. Therefore, in terms of composition, NISS and LIM are similar in composition.

[0037] Preferably, the mass fraction of the ion-free sugar aqueous solution is 10%. The sucrose solid used in the preparation of the sucrose aqueous solution is of analytically pure AR grade; the trehalose solid used in the preparation of the trehalose aqueous solution is of analytically pure AR grade. The conductivity of the deionized water used (25°C) is ≤0.055μs / cm.

[0038] Preferably, the irregular antibody is an Rh blood type anti-D antibody or an incomplete anti-E antibody.

[0039] The embodiment of the present invention also provides a method for detecting irregular antibodies, comprising the following steps: adding an ion-free sugar aqueous solution to a mixture of red blood cells expressing specific antigens and plasma to be tested, centrifuging, removing the supernatant, and then adding physiological saline to observe whether non-specific agglutination products dissipate within 1 minute. Then, the red blood cell suspension is added to a microcolumn gel anti-human globulin card and centrifuged.

[0040] Preferably, the centrifugal force during centrifugation is 1000 g.

[0041] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0042] Example

[0043] 1.1 Reagents: 1 bottle of 500g analytical grade AR grade sucrose powder reagent (Sinopharm), 4500mL deionized water (self-provided by the laboratory deionized water machine), polybrene kit (Beso), monoclonal IgG anti-D reagent (Shanghai Blood Biology), human IgG anti-E antibody plasma (remaining samples from clinical tests), mixed 6-person AB type plasma (remaining samples from clinical tests), microcolumn gel anti-human globulin reagent card (Jiangyin Libo), antibody screening red blood cells (Shanghai Blood Biology), ABO standard red blood cells (Changchun Boxun), normal saline, disposable small test tubes, micropipette and matching disposable pipette tips, 96-well plastic microplate, etc.

[0044] 1.2 Instruments: Standardized centrifuge for blood type serology (Beso), TD-2Y multifunctional centrifuge for medical testing (Zhongshan Biosciences), FYQ immune microcolumn incubator (Changchun Boyan).

[0045] 1.3 Preparation of homemade enhancer (sucrose aqueous solution):

[0046] In a suitable clean container with a lid, use 4500 mL of deionized water to dissolve 500 g of sucrose powder, i.e., prepare a 10% concentration of sucrose deionized water solution and mix it thoroughly without visible particulate matter precipitating. The solution can be stored at 4°C for a short period of time and at -20°C for long-term storage.

[0047] 1.4 Calibration of IgG anti-D and anti-E antibody titers:

[0048] In a 96-well plastic microplate, monoclonal IgG anti-D reagent and human IgG anti-E plasma were diluted in series with physiological saline. In a microcolumn gel anti-human globulin card, 0.8% cell concentration of O+ standard red blood cells and R 2 R 2 The IgG anti-D and anti-E titers were used as the calibration titers for the screening of type O antibody red blood cells (type O RhccDEE). The specific operation steps were completed in accordance with the standard operating procedures (SOP) formulated by this laboratory, and the final IgG anti-D calibration titer was 1:1024 and the IgG anti-E calibration titer was 1:8.

[0049] 1.6 Determination of IgG anti-D titer using a homemade enhancer (sucrose aqueous solution) method:

[0050] First, the irregular antibody screening test (microcolumn gel anti-human globulin method) confirmed that 6 K samples collected from the remaining samples of clinical examination 2 In EDTA-anticoagulated AB plasma, there is no additional anti-D, anti-E or other irregular antibodies that may affect the test results;

[0051] In a disposable small test tube, take 100 μL of the AB type anti-screening negative plasma to dilute the monoclonal IgG anti-D reagent (100 μL) in multiples, so that there is 100 μL of diluted plasma in each small test tube (the first tube is 100 μL of undiluted IgG anti-D reagent, the titer is 1:1), and the antibody dilutions from the second tube to the eleventh tube are 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:128, 1:256, 1:512, 1:1024, and set one tube (the twelfth tube) as a diluted plasma sample as a negative control;

[0052] Add 1 drop (about 50 μL) of 3% O+ standard red blood cells to each tube, mix manually for a few seconds, and then centrifuge in a Baso high-speed centrifuge at 3400r / min (centrifugal force 1000g) for 15s. Take out the resuspended cells and observe that the results should all be negative. This step is to exclude the interference of IgM blood type antibodies in plasma. If the result is positive, subsequent testing can still be performed, because the positive result caused by IgM antibodies will eventually be weakened by the reaction process;

[0053] Add 3 mL of the sucrose deionized water solution prepared in 1.3 to the resuspended samples, centrifuge at 3400 r / min (centrifugal force 1000 g) for 1 min, remove the samples, gently pour off the supernatant and keep some to ensure that the cell buckle at the bottom of the tube can be resuspended. After this step, gently shake each tube and observe that the red blood cell buckle is in a non-specific agglutination state. If no agglutination occurs, retest;

[0054] Immediately afterwards, add 2 drops (about 100 μL) of saline to each tube, and gently shake the test tube while observing whether the cell agglutination dissipates within 1 minute. If the agglutination does not dissipate, it is judged as positive, and the agglutination degree of "0.5+" that can be distinguished by the naked eye as a positive result is determined as the maximum dilution and recorded as the titer; repeat the above test to detect the IgG anti-E titer, the difference is that the dilution tube only needs to be diluted to 1:32 tube, and the indicator red blood cells added to each tube are R2R2 type anti-screen cells. The results are shown in Table 1 and Figure 1-2 shown.

[0055] 1.7 Determination of IgG anti-D and anti-E titers using the polybrene reagent method

[0056] Similarly, 6 portions of AB type anti-screening negative plasma were mixed as described in 1.6, and corresponding to the dilution results in 1.6, monoclonal IgG anti-D reagent and human IgG anti-E plasma were diluted in small test tubes in multiples, and then the antibody titer was detected using the polybrene kit method. The specific operation steps were carried out according to the requirements of the kit's instructions. Similarly, the positive result that can be distinguished by the naked eye - "0.5+" agglutination degree was determined as the maximum dilution and recorded as the titer. The results are shown in Table 1. Figure 1 , Figure 2 shown.

[0057] Table 1 Antibody titers

[0058] IgG antibody titer Calibration results Polybrene Sucrose aqueous solution Anti-D 1:1024 1:1024 1:1024 Anti-E 1:8 1:1 1:8

[0059] From Table 1 and Figure 1 , Figure 2 It can be concluded that in the IgG anti-D antibody titer detection, monoclonal antibody reagents (good uniformity and high antibody titer) were used, and the results of antibody titer detection by sucrose solution method and polybrene reagent method were in good consistency. However, in the IgG anti-E antibody titer detection, since the human IgG anti-E used was non-monoclonal IgG and the titer was only 1:8 (calibrated by microcolumn gel anti-human globulin method), the detection ability of polybrene reagent for IgG anti-E antibodies was lower than that of sucrose solution for IgG anti-E antibodies.

[0060] In the microcolumn gel anti-human globulin test results of a clinical patient cross-matching test, Figure 4 The agglutination degree of the secondary well (patient red blood cells + donor plasma) "+" result is Figure 4 The DAT positive result of the patient's red blood cells is consistent with the agglutination degree, which shows that the secondary side incompatibility is caused by the patient's red blood cell DAT positive, and there is a slight agglutination in the top layer of the gel in the main side hole (patient plasma + donor red blood cells). Figure 5 If the DAT result of the donor red blood cells is "-", the positive result is caused by the suspected presence of certain antibodies in the patient's plasma, and the agglutination result of this antibody and the donor red blood cells in the microcolumn gel anti-human globulin card is not obvious. Figure 4 The results of the screening of irregular antibodies in the plasma of patients were all negative, which reflected that the test was not the best method for detecting this unknown antibody. Therefore, the polybrene method and the ion-free sugar aqueous solution method were used for simultaneous detection. The results were compared as follows: Figure 3 The results of the polybrene method and the microcolumn gel anti-human globulin method were consistent, and the irregular antibody screening (including cells I, II, and III) were all negative. However, the results of the ion-free sugar aqueous solution method showed that the antibody was specific (II "0.5+", III "2+w").

[0061] When we use microcolumn gel anti-human globulin cards for direct anti-human globulin tests, we need to wash the red blood cells with saline several times to eliminate the interference of plasma proteins on the antibodies adsorbed on the red blood cells and cause false negative results. Figure 6The data showed that in a clinical cross-matching case, the DAT result of the patient's microcolumn gel anti-human globulin card method was known to be "±", and the self-control agglutination degree was weaker than the DAT result; the secondary result of the cross-matching with the blood donor by the microcolumn gel anti-human globulin card method showed "-", and after enhancement by the ion-free sugar aqueous solution method (NISS first and then card transfer), the secondary result of the cross-matching by the card method was basically consistent with the patient's DAT result.

[0062] In summary, the embodiments of the present invention provide a sucrose solution enhancer, which has simple ingredients, easy preparation, simple and easy-to-operate test steps, and uses a monoclonal IgG anti-D reagent diluted in multiple ratios and a plasma containing human IgG anti-E antibodies in a test tube-based test to make a parallel comparison with the polybrene reagent method, verifying that the reagent and method are applicable to K 2 The detection performance of IgG anti-D and anti-E blood type irregular antibodies in EDTA anticoagulated plasma samples has good consistency between the antibody titer and the titer of the polybrene method. It can replace the polybrene method for the detection of Rh blood type incomplete antibodies and can be promoted and applied in primary clinical laboratories.

[0063] The embodiments described above are part of the embodiments of the present invention, rather than all of the embodiments. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

Claims

1. Application of an ion-free sugar aqueous solution in the detection of irregular blood type antibodies.

2. The use of the non-ionized sugar aqueous solution according to claim 1 in the detection of irregular blood type antibodies, characterized in that: Application in microcolumn gel anti-human globulin detection.

3. The use of the ion-free sugar aqueous solution according to claim 1 in the detection of irregular blood type antibodies, characterized in that: The ion-free sugar aqueous solution is a sucrose aqueous solution, a trehalose aqueous solution or a mixture of the two.

4. The use of the non-ionized sugar aqueous solution according to claim 1 in the detection of irregular blood type antibodies, characterized in that: The mass fraction of the ion-free sugar aqueous solution is 10%.

5. The use of the ion-free sugar aqueous solution according to claim 3 in the detection of irregular blood type antibodies, characterized in that: The sucrose solid used in the preparation of the sucrose aqueous solution is of analytically pure AR grade; the trehalose solid used in the preparation of the trehalose aqueous solution is of analytically pure AR grade.

6. The use of the non-ionized sugar aqueous solution according to claim 3 in the detection of irregular blood type antibodies, characterized in that: The conductivity (25° C.) of the deionized water used in the preparation of the sucrose aqueous solution and the trehalose aqueous solution is ≤0.055 μs / cm.

7. A method for detecting irregular blood type antibodies, characterized in that: The following steps are involved: The test tube method is used to add the ion-free sugar aqueous solution according to any one of claims 1 to 6 to a mixture of a red blood cell suspension and plasma to be tested, centrifuge, remove the supernatant, add physiological saline, and observe whether the non-specific agglutinate dissipates within 1 minute.

8. The method for detecting irregular antibodies according to claim 7, characterized in that: The centrifugal force during centrifugation is 1000g and the centrifugation time is 1-2min.

9. The method for detecting irregular antibodies according to claim 7, characterized in that: The method also includes adding the red blood cell suspension to a microcolumn gel anti-human globulin card after adding physiological saline for solution, and centrifuging.

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