Article

[Establishment of allele specific primer polymerase chain reaction sequence-based typing and investigation of the polymorphism in exon 3 of HLA-DRB1].

Blood Center of Zhejiang Province, Key Laboratory of Blood Safety Research of Ministry of Health of China, Hangzhou, Zhejiang, P.R. China.
Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics 12/2010; 27(6):639-43. DOI:10.3760/cma.j.issn.1003-9406.2010.06.008 pp.639-43
Source: PubMed

ABSTRACT To 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.
The 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.
The 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.
The 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.

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Keywords

allele specific primer polymerase chain reaction sequence-based typing
 
Assign 3.5 SBT software
 
directions
 
exon 3
 
exon 3 sequences
 
exons 2
 
group specific primers
 
HLA-DRB1 gene
 
human leukocyte antigen( HLA)-DRB1
 
PCR-SBT
 
PCR-SBT method
 
polymorphism
 
restriction enzymes
 

Fa-Ming Zhu