Single-nucleotide polymorphism locus of ABO blood variant inducing hemolytic transfusion reaction

A technology of ABO blood type and blood transfusion reaction, which is applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial measurement/testing, etc., to achieve rapid detection and avoid the effect of acute hemolytic transfusion reaction

Active Publication Date: 2017-05-10
QINGDAO BLOOD CENT
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As long as 5-10ml of ABO-incompatible blood is transfused, obvious symptoms will appear; blood transfusion exceeding 200ml will cause serious consequences

Method used

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  • Single-nucleotide polymorphism locus of ABO blood variant inducing hemolytic transfusion reaction
  • Single-nucleotide polymorphism locus of ABO blood variant inducing hemolytic transfusion reaction
  • Single-nucleotide polymorphism locus of ABO blood variant inducing hemolytic transfusion reaction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Confirmation of the mutation site of type A variant from the gene detection of the proband of ABO blood type A variant gene.

[0027] 1. Extraction of peripheral blood genomic DNA:

[0028] In line with the relevant national policies and regulations, and on the basis of the consent of the sampling subjects, draw 2-5ml of peripheral venous blood from members of the ABO blood type A variant gene family and put it into EDTA-Na 2 In anticoagulant tubes, freeze at -80°C for later use; DNA extraction uses the DNA extraction kit from LIFE Company, and the specific operation process is as follows:

[0029] After thawing the frozen EDTA anticoagulated blood at room temperature, take 500ul and put it in a centrifuge tube, add an equal volume of erythrocyte lysate (pH8.0), vortex and mix for 5 minutes, centrifuge at 12000rpm for 5 minutes, and discard the supernatant. Add 500ul of red blood cell lysate repeatedly, vortex and mix for 5 minutes, centrifuge at 12000rpm fo...

Embodiment 2

[0041] Verification of Example 2 SNP site

[0042] 3. Perform haplotype sequencing on exon 7 of the ABO gene to determine the mutation site of the A variant gene in the proband

[0043] Primers for PCR amplification of target fragments: ABO-E7F: 5'-ATCCGCCTGCCTTGCAGATA-3'; ABO-E7R: 5'-AGCCTAGGCTTCAGTTACTCACAACA-3'.

[0044]The PCR amplification primers may also select other primer pairs capable of amplifying the above-mentioned SNP sites.

[0045] Reaction conditions and reaction system for PCR amplification of target fragments:

[0046] (1) PCR reaction conditions: pre-denaturation at 95°C for 5 minutes; 40 cycles of 95°C for 30 seconds, 60°C for 30 seconds, and 72°C for 50 seconds; extension at 72°C for 5 minutes, and incubation of the amplified product at 10°C.

[0047] (2) Reaction system: (TAKARA LA-Taq polymerase)

[0048]

[0049] The reaction system was used to carry out the amplification reaction between the genomic DNA template of each family member and the pri...

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Abstract

The invention aims to provide a single-nucleotide polymorphism locus for detecting A-type variant, inducing acute hemolytic transfusion reaction, of an ABO blood-group system. The single-nucleotide polymorphism locus refers to the 689th basic group counted from an initiation codon in an A-type variant gene coding area. The single-nucleotide polymorphism locus has the advantages that new application of an A-type variant gene is provided, an effective way to fast gene diagnosis, gene screening and genetic counselling for induced acute intravascular hemolytic transfusion reaction is provided, and according to application effects, the single-nucleotide polymorphism locus of the gene and a detect primer can be applied to fast A-type variant gene mutation locus detection for peripheral blood of a clinical patient.

Description

technical field [0001] The invention belongs to the technical field of gene diagnosis products, and in particular relates to a SNP site for detecting acute or delayed hemolytic transfusion reactions caused by type A variant blood type genes in the ABO blood type system. Background technique [0002] The normal ABO blood type is divided into four types: A, B, AB, and O. However, in the case of ABO blood type gene mutations such as SNP, deletion, insertion, duplication, etc., the ABO blood type erythrocyte surface antigen and plasma For example, in the ABO blood group system found in the present invention, the A-type variant blood type gene, the A antigen on the red blood cell is weaker than that of the normal A-type person, but there is an anti-A1 cell antibody in the plasma, once infused Type A blood is prone to transfusion hemolysis. [0003] Hemolytic transfusion reaction (Hemolytic Transfusion Reaction; HTR) is divided into acute and delayed hemolytic transfusion reactio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 冯智慧马文慧胡彬韩斌
Owner QINGDAO BLOOD CENT
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