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Sorting type RBC (Red Blood Cell) blood type irregular antibody detection kit based on solid phase agglutination technology and preparation method thereof

A technology for antibody detection and erythrocytes, which is applied in biological testing, measuring devices, and analytical materials, can solve the problems of high cost, short storage period of erythrocytes, cumbersome operation, etc., and achieve the effect of convenient storage and improved coating efficiency

Pending 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 sorting-type red blood cell blood type irregular antibody detection kit based on solid-phase agglutination technology, which can solve the problems of short red blood cell storage period, missed antibody detection, cumbersome operation, and high cost in existing detection methods. higher question

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  • Sorting type RBC (Red Blood Cell) blood type irregular antibody detection kit based on solid phase agglutination technology and preparation method thereof
  • Sorting type RBC (Red Blood Cell) blood type irregular antibody detection kit based on solid phase agglutination technology and preparation method thereof
  • Sorting type RBC (Red Blood Cell) blood type irregular antibody detection kit based on solid phase agglutination technology and preparation method thereof

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Effect test

Embodiment 1

[0042] The preparation of embodiment 1 sorting type erythrocyte blood group irregularity 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 split-type solid-phase agglutination reaction microplates

[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) Prepare sieve-resistant cells numbered Ⅰc, Ⅱc, and Ⅲc...

Embodiment 2

[0056] Example 2 Separation-type solid-phase agglutination reaction microplate before and after freeze-drying Erythrocyte membrane antigenicity test

[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 it as -D, -E, -Jk from top to bottom a ,-Jk b 、-Fy a...

Embodiment 3

[0064] Example 3 The detection kit for the detection of irregular red blood cell blood group antibodies in the detection of high-frequency antibodies in the Chinese population. Comparison of anti-Mur and conventional anti-screening 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 1st to 3rd wells from top to bottom (i.e. Ic, IIc, and IIIc anti-sieve cells) as -D, -E, -Mur, the 4th to 6th wells (i.e. Ic, Ⅱc, and Ⅲc anti-sieve cells) are also marked as -D, -E, -Mur as the re-examination wells of the 1st to 3rd wells, and...

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Abstract

The invention relates to a sorting type RBC (Red Blood Cell) blood type irregular antibody detection kit based on solid phase agglutination technology and a preparation method thereof, and belongs tothe technical field of preparation of kits. The kit comprises (1) a sorting type solid phase agglutination reaction micro-plate; (2) a low ionic strength solution; (3) an indicator system; (4) a negative control serum; and (5) a positive control serum. The preparation method comprises the following steps of (1) preparation of the sorting type solid phase agglutination reaction micro-plate; (2) preparation of the low ionic strength solution; (3) preparation of the indicator system; (4) preparation of the negative control serum; and (5) preparation of the positive control serum. Through the innovative two-step indicator system using high specific adsorption of monoclonal antibodies to RBCs, using the freeze-drying preservation technology and using high sensitivity, problems such as low sensitivity, difficult storage and easy miss detection of RBCs in the current clinical irregular antibody detection methods are practically solved, and a cost-effective detection kit for clinical first-line detection work is provided.

Description

technical field [0001] The invention relates to a sorting-type erythrocyte blood type irregular antibody detection kit and a preparation method based on solid-phase agglutination technology, 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, once t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/80G01N33/543
CPCG01N33/80G01N33/54306
Inventor 骆宏罗广平
Owner 广州血液中心
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