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A kind of pool-checking red blood cell blood group irregular antibody detection kit based on solid-phase agglutination technology and preparation method

A technology for antibody detection and red blood cells, which is applied in biological testing, material inspection products, and disease resistance to media transmission. It can solve the problems of short storage period of red blood cells, missed antibody detection, and cumbersome operation, so as to improve detection sensitivity and avoid missed detection. , The effect of simple operation process

Active Publication Date: 2022-07-12
广州血液中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] An object of the present invention is to provide a detection kit for irregular red blood cell blood group antibodies based on solid-phase agglutination technology, which can solve the problems of short storage period of red blood cells, missed detection of antibodies, cumbersome operation and high cost in existing detection methods. higher question

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  • A kind of pool-checking red blood cell blood group irregular antibody detection kit based on solid-phase agglutination technology and preparation method
  • A kind of pool-checking red blood cell blood group irregular antibody detection kit based on solid-phase agglutination technology and preparation method
  • A kind of pool-checking red blood cell blood group irregular antibody detection kit based on solid-phase agglutination technology and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Preparation of Hui-check-type erythrocyte blood group irregular antibody detection kit

[0043] 1. Formation of irregular antibody screening lineages: Monoclonal antibodies are used to screen O-type red blood cells. According to the principle of antigen complementation, an antibody-screening cell line composed of 3 O-type red blood cells is established. The specific antigen typing spectrum is shown in Table 1;

[0044] 2. Preparation of the pooled solid-phase agglutination reaction microplate

[0045] (1) Dilute the anti-erythrocyte monoclonal antibody with carbonate coating solution to 100ug / mL, take out a blank 96-well reaction microplate, add 100 μL to each well, coat at room temperature for 16 hours, and dry the liquid in the microwell After washing with purified water for 3 times, the residue in the micropores was dried with absorbent paper for the last time;

[0046] (2) The anti-sieve cells numbered Ⅰc, Пc, and Шc were washed three times with normal sa...

Embodiment 2

[0056] Example 2 Erythrocyte membrane antigenicity test before and after lyophilization of solid-phase agglutination reaction microplates

[0057] Select D, E, Jk with dose effect a , Jk b , Fy a The antigen is used as the target antigen, and the kit of the present invention is used to detect the human plasma containing the above-mentioned antibodies to evaluate whether the antigenicity of the red blood cell membrane changes after freeze-drying. The experimental results show that the detection results of the above antibodies on the micro-strips before and after freeze-drying are consistent, all of which are 4+, indicating that the antigenicity of the red blood cell membrane does not change before and after freeze-drying. The results are shown in image 3 . The specific operation method is as follows:

[0058] 1. Take out one microplate strip from the kit prepared in Example 1, equilibrate to room temperature, and mark the 1st to 8th wells as -D, -E, -Jk from top to bottom....

Embodiment 3

[0064] Example 3 Comparison of anti-Mur and conventional anti-screening cells for the detection of high-frequency antibodies in Chinese population

[0065] The high-frequency antibody anti-Mur in the Chinese population was used as the target to detect antibodies, and compared with conventional anti-screening cells. As a result, the kit of the present invention can better detect anti-Mur antibodies, while conventional anti-sieve cells cannot be detected, see Figure 4 . The specific operation method is as follows:

[0066] 1. After taking out one microplate strip from the kit prepared in Example 1, equilibrate to room temperature, and label the 1st to 8th wells as Normal (normal plasma), -Mur, -D, Normal ( normal plasma), -Mur, -D, Neg (negative control), Pos (positive control);

[0067] 2. Add 2 drops (100 μL) of low-ion solution to each of the above wells, and add 1 drop (50 μL) of the test specimen and negative and positive control serum corresponding to the label to the ...

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Abstract

The present invention relates to a detection kit based on solid-phase agglutination technology and a preparation method for the detection kit of erythrocyte irregular blood group antibodies, belonging to the technical field of kit preparation. (2) Low ionic strength solution; (3) Indicator system; (4) Negative control serum; (5) Positive control serum; the preparation method includes: (1) Preparation of pool-detection solid phase agglutination reaction microplate; (2) ) Preparation of low ionic strength solution; (3) Preparation of indicator system; (4) Preparation of negative control serum; (5) Preparation of positive control serum; The freeze-drying preservation technology and the high-sensitivity two-step indicator system can effectively solve the problems of low sensitivity, difficult to preserve red blood cells, and easy to miss detection in the current clinical irregular antibody detection methods, and provide a cost-effective detection for clinical first-line detection work. Reagent test kit.

Description

technical field [0001] The invention relates to a collection-type erythrocyte blood group irregular antibody detection kit based on solid-phase agglutination technology and a preparation method, and belongs to the technical field of kit preparation. Background technique [0002] Irregular antibodies of the erythrocyte blood group system refer to blood group antibodies other than anti-A and anti-B in the human body. In addition to ABO antigens in human erythrocytes, common antigens include D, C, E, c, e, and JK. a , JK b , M, N, S, s, Fy a , Fy b , Lu b Wait. Correspondingly, antibodies raised against these antigens belong to the category of irregular antibodies. [0003] Irregular antibodies are mostly immune antibodies, which are produced mainly by blood transfusion, pregnancy, and exposure to natural immune antigens. These antibodies are mainly of the IgG type. For the blood recipient, when there are irregular antibodies in the body, once the red blood cells positi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/80
CPCG01N33/80Y02A50/30
Inventor 骆宏罗广平王嘉励魏玲
Owner 广州血液中心
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