A group-specific primer PCR-SBT method and reagents for hla-dqb1 genotyping

A genotyping and specific technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve the problems of DQB1*02 allele missed detection, allele missed detection, etc., to reduce rejection and improve accuracy Sexuality, the effect of improving the success rate and survival rate

CN104894230BActive Publication Date: 2018-06-12浙江省血液中心
1 Cites 0 Cited by

Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2018-06-12

Smart Images

  • Figure 1
    Figure 1
  • Figure 2
    Figure 2
  • Figure 3
    Figure 3
Patent Text Reader

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.
Need to check novelty before this filing date? Find Prior Art

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...

Examples

Embodiment Construction

[0052] The content of the present invention will be described in further detail below in conjunction with the examples.

[0053] This implementation specifically takes one specimen for HLA-DQB1 genotyping as an example to describe the content of the present invention in detail. A group-specific primer PCR-SBT method for HLA-DQB1 genotyping used in the present invention specifically includes the following steps:

[0054] 1. Prepare human genomic DNA as a template for PCR amplification in subsequent steps.

[0055] 200 µl of whole blood to be tested was taken, and genomic DNA was extracted according to the instructions of the QuickGene DNA whole blood kit S kit, and the concentration and purity of the genome were determined by a spectrophotometer.

[0056] 2. Synthesize 6 pairs of amplification primers and 4 sequencing primers. For the specific sequence, please refer to the sequence in the content of the invention mentioned above, which will not be repeated. Dilute the amplifica...