title: [Establishment of allele specific primer polymerase chain reaction sequence-based typing and investigation of the polymorphism in exon 3 of HLA-DRB1].
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Blood Center of Zhejiang Province, Key Laboratory of Blood Safety Research of Ministry of Health of China, Hangzhou, Zhejiang, P.R. China. zfm00@hotmail.com
authors.forename: Fa-ming
authors.initials: FM
authors.lastname: Zhu
authors.forename: Yan-min
authors.initials: YM
authors.lastname: He
authors.forename: Su-dan
authors.initials: SD
authors.lastname: Tao
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Jun-jun
authors.initials: JJ
authors.lastname: He
authors.forename: Hang-jun
authors.initials: HJ
authors.lastname: Lv
authors.forename: Li-xing
authors.initials: LX
authors.lastname: Yan
mesh_terms: D000483:Alleles; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics*; D019143:Evolution, Molecular; D005091:Exons / Q000235:genetics*; D005838:Genotype; D006684:HLA-DR Antigens / Q000235:genetics*; D059811:HLA-DRB1 Chains; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic / Q000235:genetics*
chemical_list: D017931:DNA Primers; D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains