title: [Establishment of a polymerase chain reaction sequencing based typing method for HLA-DPB1 exons 2 and 3 and investigation of their polymorphisms].
abstract: OBJECTIVE
  To establish a polymerase chain reaction sequencing-based typing (PCR SBT) method for HLA-DPB1 exons 2 and 3, and to analyze their polymorphisms.
  
  
  METHODS
  Based on the sequences of HLA-DPB1 loci, locus-specific primers were designed and applied to amplify the target sequences encompassing the entire exons 2 and 3 of HLA-DPB1. The amplification products were digested by enzymes and directly sequenced in both directions. The genotype was assigned by Assign 3.5+ SBT software.
  
  
  RESULTS
  Specific target fragment was obtained with the PCR amplification, and good quality electropherogram was derived by direct sequencing. Among 242 individuals from Zhejiang Han population, 18 HLA-DPB1 alleles were detected. Alleles with a frequency of > 0.05 have included DPB1*05:01:01/135:01 (0.4112), DPB1*02:01:02 (0.1901), DPB1*04:01:01 (0.1136) and DPB1*02:02 (0.0620). A novel HLA-DPB1*168:01 allele has also been identified. Nine polymorphism sites were founded in the exon 3 region, which included a new SNP site 517 A>T.
  
  
  CONCLUSIONS
  The PCR-SBT method for exons 2 and 3 of HLA-DPB1 is reliable, which allowed detection of polymorphisms in exon 3 of the HLA-DPB1 gene.
authors: Blood Center of Zhejiang Province, Hangzhou, Zhejiang 310006, P. R. China. Email:zfm00@hotmail.com.
authors: Yanmin
authors: Y
authors: He
authors: Sudan
authors: S
authors: Tao
authors: Wei
authors: W
authors: Zhang
authors: Wei
authors: W
authors: Wang
authors: Ji
authors: J
authors: He
authors: Faming
authors: F
authors: Zhu
authors: Hangjun
authors: H
authors: Lyu
mesh_terms: D000483:Alleles; D005091:Exons; D059868:HLA-DP beta-Chains / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic
chemical_list: D059868:HLA-DP beta-Chains; C071170:HLA-DPB1 antigen