Group-specific primer PCR-SBT method and reagent based on HLA-DQB1 genetic typing

Abstract

The invention provides a group-specific primer PCR-SBT reagent based on HLA-DQB1 genetic typing. The group-specific primer PCR-SBT reagent is composed of six pairs of group-specific primers for amplification and four oligonucleotides sequencing primers for sequencing analysis. The invention furthermore provides a group-specific primer PCR-SBT method based on HLA-DQB1 genetic typing adopting the reagent. The reagent and method can be used as an independent widely-applied identification method. The accurate typing identification problem of HLA-DQB1 sites can be successfully solved. Accuracy of HLA matching of hematopoietic stem cell transplantation donors and recipients can be improved. Therefore, more proper transplantation donors are selected, and rejection reactions in the transplantation process are reduced. Great significance in furthermore improving the success rate and survival rate of organ transplantation is achieved.

Description

technical field [0001] The invention relates to a genotyping detection method, in particular to a molecular biology detection method for HLA-DQB1 genotyping, and also relates to reagents and related experimental parameters used in the method. 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. Accurate HLA 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-DQB1 belongs to the classic HLA-II gene a...

AI Technical Summary

Problems solved by technology

Foreign scholars have reported that the DQB1 gene is prone to allele under-detection in the case of heterozygosity such as DQB1*02, DQB*03, and DQBI*04

Domestic studies have also reported that when DQB1*06 and DQB1*02 are heterozygous, the DQB*06 allele is preferentially amplified, and when DQB1*03 and DQB1*02 are heterozygous, the DQB1*03 allele is preferentially amplified, resulting in DQB1* 02 allele missed detection

In our previous research, we also found that although the existing commercial kits can simultaneously detect exon 2 and exon 3 of HLA-DQB1, there are similar cases of allele missing

Therefore, there are certain defects in the existing methods. Establishing a PCR-SBT method for a single allele of HLA-DQB1 can ensure the effective amplification of different alleles, which is of great significance for providing accurate HLA-DQB1 typing results

Images