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SNP site for AB variant type (AwB) capable of triggering hemolytic transfusion reaction (HTR)

A blood transfusion reaction, AB-type technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, microbial determination / inspection, etc., to achieve rapid detection and avoid the effect of acute hemolytic transfusion reaction

Inactive Publication Date: 2019-03-26
QINGDAO BLOOD CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Liu Jianling et al. reported that a pair of twins in the family were both of B(A) blood type, and they were diagnosed with xeroderma pigmentosa after birth. There is no theoretical basis to find whether it is related to B(A) blood type.

Method used

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  • SNP site for AB variant type (AwB) capable of triggering hemolytic transfusion reaction (HTR)
  • SNP site for AB variant type (AwB) capable of triggering hemolytic transfusion reaction (HTR)
  • SNP site for AB variant type (AwB) capable of triggering hemolytic transfusion reaction (HTR)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Confirmation of the mutation site of the AB variant (AwB) from the genetic testing of the proband of the ABO blood type AwB variant gene.

[0028] 1. Extraction of peripheral blood genomic DNA:

[0029] In compliance 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 AB 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:

[0030] 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, centr...

Embodiment 2

[0041] Verification of Example 2 SNP site

[0042] 3. Perform haplotype sequencing on exons 6, 7 and intron 6 of the ABO gene to determine the mutation site of the AB variant gene in the proband

[0043] Primers for PCR amplification of the target fragment: ABO-E6mo: 5'-CGGGATCCATGTGGGTGGCACCCTGCCA-3'; ABO-E7mo: 5'-GGGCCTAGGCTTCAGTTACTC-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...

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Abstract

The invention aims to provide an SNP site for detecting the AB variant type (AwB) capable of triggering an acute or delayed hemolytic transfusion reaction (HTR). The SNP site is the 803 basic group starting from initiation codon in an ABO blood type gene encoding region, and is mutation with G being greater than C. The invention provides new use of AwB type gene detection of the ABO blood type, thus effective genetic diagnosis, prenatal genetic screening and genetic counseling paths for effectively avoiding the acute HTR are provided, the application effects show that the SNP site of the gene and detecting primers can be effectively used for peripheral blood of clinical patients for quickly detecting the gene mutation site of the ABO blood type AwB type.

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 AB variants (AwB) that trigger acute or delayed hemolytic transfusion reactions. 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 AwB blood type in the ABO blood type system found in the present invention, 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 the AB type person is infused Blood, prone to transfusion hemolysis. [0003] Hemolytic transfusion reaction (Hemolytic Transfusion Reaction; HTR) is divided into acute and delayed hemolytic transfusion reaction. Blood transfusion with incompatible ABO...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 刘丽逄淑涛韩斌冯智慧
Owner QINGDAO BLOOD CENT
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