HLA-C allele sequencing typing method

Abstract

The invention discloses an HLA-C allele sequencing typing method. The method comprises the following steps: PCR (polymerase chain reaction) amplification, PCR product electrophoresis detection, PCR product purification, sequencing reaction, sequencing product ethanol purification, device sequencing and data analysis. The method can implement simultaneous amplification on the 1-4 exons of the HLA-C site by using one primer pair, can be used for sequencing, and is simple to operate and easy to implement.

Description

technical field [0001] The invention belongs to the field of gene sequencing, in particular to a method for sequencing and typing HLA-C alleles. Background technique [0002] Human leukocyte antigen (HLA) is located in the 6p2113 region of the short arm of chromosome 6, which is the most complex genetic polymorphism system in the human genome. HLA-C belongs to the classic HLA class I gene, which is located between HLA-A and B sites. HLA-C molecules not only present endogenous polypeptides to CD8+ T cells, induce specific cell killing effects, but also act as natural killer cells. Immunoglobulin-like receptor (KIR) ligands, the combination of the two can initiate inhibitory signals, and play an important role in transplantation immunity, tumor immunity and anti-infection immunity. The expression level of HLA-C antigen on the cell surface is low, and fewer antigens can be detected by serological methods. In the past, it was believed that the polymorphism of HLA-C molecules wa...

AI Technical Summary

Problems solved by technology

[0003] Delfino and Dormoy et al.'s HLA-C allele sequencing and typing methods are respectively in HLA-C sequence based typing: nucleotide analysis from exon 1through exon 8. Identification of a new allele: Cw*0718 (2003Tissue Antigens 62 (5): 418- 425) and Mono-allelic amplification of exons 2-4 using allele group-specific primers for sequence-based typing(SBT) of the HLA-A,-B and-C genes: preparation and validation of ready-to-use pre-SBT mini- Kits (2003, Tissue Antigens 62(3):201-216.) made a clear elaboration, but the method adopted by Delfino et al., its 1-3 exons and 4-8 exons are amplified separately, and the amplified The amplification conditions are different, so exons 1-4 cannot be amplified and detected under the same conditions; while the method of Dormoy et al. amplifies exons 2-4, and cannot amplify all HLA-C alleles with a single primer Gene

Images