Reagent for PCR-SBT method for HLA genotyping

A PCR-SBT, genotyping technology, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as affecting clinical work and alleles cannot be specified.
CN101928776BActive Publication Date: 2012-10-10浙江省血液中心
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
浙江省血液中心
Publication Date
2012-10-10

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Abstract

The invention provides a PCR-SBT method for HLA genotyping. Exons 1, 2, 3, 4, 5, 6, 7 of an HLA-A locus, an HLA-B locus and an HLA-C locus are subjected to genotyping. The method comprises the following steps of: (1) preparing human genomic DNA; (2) designing an amplification primer; (3) performing double digestion purification on an amplification product; (4) designing a sequencing primer and performing sequencing PCR on a purification product; (5) purifying a sequencing product by a sodium acetate-ethanol precipitation method and performing capillary electrophoresis sequencing; and (6) performing software analysis on the acquired sequence to determine genotypes thereof. The exons 1, 2, 3, 4, 5, 6, 7 of the HLA-A, B, C loci are subjected to full-length sequence sequencing, so that the full length of the related exons of the HLA-A, B, C loci and part of intron oligonucleotide sequences are obtained and the HLA genotyping is accurately performed.
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Description

technical field

[0001] The invention relates to a genotyping detection method and a reagent thereof, in particular to a reagent used in a molecular biology detection method for genotyping of exons 1-7 of HLA-A, B, and C sites. Background technique

[0002] The human leukocyte antigen (HLA) gene is located in the 21.3 region of the short arm of chromosome 6. It is the main gene system that regulates the specific immune response of the human body and is the most polymorphic genetic system known so far. HLA antigens are closely related to the rejection of allogeneic organ transplantation, and the survival of the graft after organ transplantation largely depends on whether the HLA types of the donor and the recipient are compatible. HLA locus typing is of great significance for selecting suitable donors, reducing the incidence of graft-versus-host disease (GVHD), and improving the survival rate of grafts.

[0003] HLA-A, -B, -C locus genes are classic HLA class I genes with hig...

Claims

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