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Human ABO blood type reverse typing colloidal gold test strip as well as preparation method and application thereof

A technology for colloidal gold test strips and ABO blood type, applied in the field of blood type detection, can solve the problems of high detection cost, high technical requirements for blood coagulation test, prone to errors, etc., and achieves the effects of strong specificity, low technical requirements and high sensitivity

Inactive Publication Date: 2017-05-31
何伟
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The hemagglutination test has the disadvantages of high technical requirements, strong subjectivity, error-prone, and high cost. At the same time, the standard A, B, and O red blood cells used have a short shelf life and are prone to microbial contamination and hemolysis.
The micro-column gel method requires a special gel card and a special centrifuge at the same time, and the detection cost is relatively high

Method used

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  • Human ABO blood type reverse typing colloidal gold test strip as well as preparation method and application thereof
  • Human ABO blood type reverse typing colloidal gold test strip as well as preparation method and application thereof
  • Human ABO blood type reverse typing colloidal gold test strip as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1 A kind of human ABO blood group reverse stereotyping colloidal gold test strip

[0046] like figure 1 As shown, a human ABO blood group reverse typing colloidal gold test strip is composed of a bottom plate (1), a sample pad (2), a colloidal gold pad (3), a nitrocellulose membrane (5) and an absorbent paper (4).

[0047] A layer of nitrocellulose membrane (5) is laid in the middle of the bottom plate (1), and the detection area of ​​the nitrocellulose membrane (5) is provided with an antibody A color band (5-1) and an antibody B color band (5-2). and quality control strips (5-3). The human blood group anti-A antibody is adsorbed on the A antibody color band (5-1); the human blood group anti-B antibody is adsorbed on the B antibody color band (5-2); the quality control band (5-2) -3) Adsorb human blood group anti-A antibody and human blood group anti-B antibody double antibody.

[0048] A sample pad (2) and a colloidal gold pad (3) are arranged at the sam...

Embodiment 2

[0053] Embodiment 2 A kind of preparation method of human ABO blood group reverse stereotyping colloidal gold test strip

[0054] Methods as below:

[0055] Trizol plus, PrimeScriptTM RT-PCR kit, Prime STAR PCR Mix, DL2000Marker, DL5000 Marker, ligation mix ligation kit, BamHI, XhoI restriction enzymes, IPTG and other reagents were purchased from Bao Bioengineering (Dalian) Co., Ltd.

[0056] Plasmid extraction kit, DNA gel recovery kit, etc. were purchased from OMEGA.

[0057] (1) Genetically recombinant human blood group A antigen protein

[0058] 1. Extraction of total DNA from human whole blood

[0059] Four cases of Chinese Han nationality voluntary blood donors were collected. According to the operating procedures of the blood bank, venous whole blood was collected, and the rapid salting-out method was used to extract genomic DNA from the buffy coat layer of the whole blood as a DNA template.

[0060] 2. PCR amplification

[0061] 1) Primer design

[0062] According...

Embodiment 3

[0143] Example 3 Application of a Colloidal Gold Test Strip for Anti-typing of Human ABO Blood Type

[0144] (1) The method is as follows

[0145] Adopt the colloidal gold test strip of people's ABO blood group reverse stereotyping prepared in embodiment 2.

[0146] Drop 50ul of the serum sample to be tested on the sample pad 1 and wait for 3 to 5 minutes. With capillary action, the blood sample moves forward and reacts with the human blood group A antigen protein or human blood group B antigen protein on the colloidal gold pad, continue Go ahead, judge the blood type of the blood sample to be tested according to the color development on the A antibody color band (5-1) and / or B antibody color band (5-2), the control line must develop color, otherwise the detection invalidated.

[0147] (2) Judgment of results

[0148] If the color band of the B antibody develops color, it is type A blood; if the color band of the A antibody develops color, it is type B blood; If the color ...

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Abstract

The invention relates to a human ABO blood type reverse typing colloidal gold test strip as well as a preparation method and application thereof. The human ABO blood type reverse typing colloidal gold test strip is composed of a base plate, a sample pad, a colloidal gold pad, a nitrocellulose membrane and absorbent paper. When a to-be-detected sample is dripped into a sample adding area, specific binding occurs between type A and B antibodies in the samples and colloidal gold labeled antigens A and B on the test strip, the compound moves forwards along the nitrocellulose membrane under the siphon action and is captured by antibodies in a detection area when being moved into the detection area, an antibody-antigen-antibody-colloidal gold double-antibody sandwich compound is formed and accumulated on a detection line to present a visible developing reaction. If the to-be-detected antibody does not exist in the sample, binding does not occur, that is, developing is avoided. The test strip disclosed by the invention has the advantages of being high in sensitivity, convenient to use, objective in results, low in cost, convenient to preserve and transport, and the like.

Description

technical field [0001] The invention belongs to the field of blood group detection, in particular to a colloidal gold reagent strip for reverse typing of human ABO blood group and a preparation method thereof. Background technique [0002] The ABO blood group system is a blood type system based on the presence or absence of specific antigens A and B on the surface of red blood cells. It divides blood types into O, A, B, and AB blood types. Type A red blood cells contain A antigens and anti-B antibodies in serum; type B red blood cells contain B antigens and anti-A antibodies in serum; type O red blood cells have no A and B antigens, and serum contains anti-A and anti-A antibodies. Anti-B antibodies; type AB red blood cells contain two antigens, A and B, but the serum does not contain anti-A and anti-B antibodies. A antigen and anti-A antibody, B antigen and anti-B antibody will have an immune binding reaction, causing red blood cells to aggregate. [0003] The ABO blood g...

Claims

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Application Information

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IPC IPC(8): G01N33/80
CPCG01N33/80
Inventor 海洋赵洪礼康慧颖孙雪薇魏冉杨振王昆何伟
Owner 何伟
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