Establishment of allele specific primer polymerase chain reaction sequence-based typing and investigation of the polymorphism in exon 3 of HLA-DRB1.
- Author:
Fa-ming ZHU
1
;
Yan-min HE
;
Su-dan TAO
;
Wei ZHANG
;
Wei WANG
;
Jun-jun HE
;
Hang-jun LV
;
Li-xing YAN
Author Information
- Publication Type:Journal Article
- MeSH: Alleles; Base Sequence; DNA Primers; genetics; Evolution, Molecular; Exons; genetics; Genotype; HLA-DR Antigens; genetics; HLA-DRB1 Chains; Humans; Molecular Sequence Data; Phylogeny; Polymerase Chain Reaction; methods; Polymorphism, Genetic; genetics
- From: Chinese Journal of Medical Genetics 2010;27(6):639-643
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish the allele specific primer polymerase chain reaction sequence-based typing (PCR-SBT) and investigate the polymorphism of exon 3 of human leukocyte antigen( HLA)-DRB1.
METHODSThe fragment containing exons 2 and 3 of HLA-DRB1 gene was amplified by group specific primers. The amplified products were digested by restriction enzymes and directly sequenced in both directions. The genotype was assigned by using Assign 3.5 SBT software.
RESULTSThe exon 3 sequences of HLA-DRB1*08:09 and HLA-DRB1*12:02:01 were identified for the first time. There were 27 polymorphic sites in exon 3 among the twenty-five HLA-DRB1 alleles, which was 9.56% of all nucleotides of exon 3. The method could discriminate the HLA-DRB1*14:01:01/14:54 ambiguous samples, and the HLA-DRB1*14:01:01 was identified in the Chinese population.
CONCLUSIONThe PCR-SBT method for exon 3 of HLA-DRB1 from the present study was reliable and there were polymorphisms in exon 3 of HLA-DRB1.
