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Convergent detection type erythrocyte blood type irregular antibody detection kit based on solid-phase agglutination technology and preparation method thereof

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

Active Publication Date: 2019-08-27
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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  • Convergent detection type erythrocyte blood type irregular antibody detection kit based on solid-phase agglutination technology and preparation method thereof
  • Convergent detection type erythrocyte blood type irregular antibody detection kit based on solid-phase agglutination technology and preparation method thereof
  • Convergent detection type erythrocyte blood type irregular antibody detection kit based on solid-phase agglutination technology and preparation method thereof

Examples

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Effect test

Embodiment 1

[0042] Example 1 The preparation of the collection type erythrocyte blood type irregular antibody detection kit

[0043] 1. Establishing an irregular antibody screening spectrum cell line Monoclonal antibodies were used to screen type O red blood cells. According to the principle of antigen complementarity, an antibody screening cell line composed of three type O red blood cells was established. The specific antigen typing spectrum is shown in Table 1;

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

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

[0046] (2) Wash the anti-sieve cells numbered Ic, Пc, and Шc three t...

Embodiment 2

[0056] Example 2 Test of Erythrocyte Membrane Antigenicity Before and After Freeze-drying of Solid-Phase Agglutination Reaction Microplate

[0057] Select D, E, Jk with dose effect a , Jk b 、Fy a The antigen is used as the target antigen, and the kit in the present invention is used to detect the human plasma containing the above antibody to evaluate whether the antigenicity of the erythrocyte membrane changes after freeze-drying. The experimental results show that the detection results of the above-mentioned antibodies on the microplate strips before and after freeze-drying are consistent, both are 4+, indicating that the antigenicity of the red blood cell membrane has not changed 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 wells 1-8 as -D, -E, and -Jk from top to bottom a ,-Jk b 、-Fy...

Embodiment 3

[0064] Example 3 Huijian Type Irregular Red Blood Cell Antibody Detection Kit Detects High-Frequency Anti-Mur Antibodies in the Chinese Population Compared with Conventional Anti-Screen Cells

[0065] The high-frequency antibody anti-Mur in the Chinese population was used as the target to detect antibodies, and compared with conventional anti-screen cells. As a result, the kit of the present invention can detect the anti-Mur antibody preferably, while the conventional anti-screen cells fail to detect, 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 mark the 1-8 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 specimen to be tested corresponding to the labe...

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Abstract

The invention relates to a convergent detection type erythrocyte blood type irregular antibody detection kit based on a solid-phase agglutination technology and a preparation method thereof, and belongs to the technical field of kit preparation. The kit comprises (1) a convergent detection type solid-phase agglutination reaction micro-plate; (2) a low-ion-strength solution; (3) an indication system; (4) negative control serum; and (5) positive control serum. The preparation method comprises the following steps of (1) preparing the convergent detection type solid-phase agglutination reaction micro-plate; (2) preparing the low-ion-strength solution; (3) preparing the indication system; (4) preparing the negative control serum; and (5) preparing the positive control serum; and through innovative utilization of high specificity adsorption of the erythrocyte by virtue of a monoclonal antibody, a freeze-drying preservation technology and a high-sensitivity two-step method indication system,the problems of low sensitivity, difficulty in storing the erythrocyte, easiness in leak detection and the like in an existing clinically irregular antibody detection method are practically solved, and the high-cost-performance detection kit is provided for the clinical front-line detection work.

Description

technical field [0001] The invention relates to a collection-type red blood cell blood type irregular antibody detection kit based on solid-phase agglutination technology and a preparation method, belonging 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 red blood cells, common antigens include D, C, E, c, e, 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] Most of the irregular antibodies are immune antibodies, and their production methods mainly include: blood transfusion, pregnancy, exposure to natural immune antigen substances, etc. These antibodies are predominantly of the IgG type. For blood recipients, when there are irregular antibodies in the body, ...

Claims

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Application Information

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Patent Type & Authority Applications(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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