title: Rapid and reliable genotyping of HLA-B*27 in the Chinese Han population using a duplex real-time TaqMan PCR assay.
abstract: OBJECTIVE
  To develop a duplex real-time TaqMan PCR assay for genotyping HLA-B*27 in the Chinese Han population.
  
  
  METHODS
  A standard curve was constituted to deduce amplification efficiency, dynamic range and detection limit of the duplex real-time TaqMan PCR assay, whereas PCR-SBT (PCR with sequence-based typing) was used to evaluate the accuracy of the assay.
  
  
  RESULTS
  A linear standard curve for determining HLA-B*27 was obtained within the range of 10(1)-10(9) copies per reaction with the correlation coefficient of 0.99 and amplification efficiency of 98.30%. The detection limit was 3.09 copies per reaction. Complete concordance was found between the results obtained by the duplex real-time TaqMan PCR assay and PCR-SBT. Fifty-nine of the 178 genomic samples were HLA-B*27 positive and the other 119 were HLA-B*27 negative.
  
  
  CONCLUSIONS
  The duplex real-time TaqMan PCR approach appears to be a reliable, sensitive, rapid and high-throughput method to genotype HLA-B*27 in the Chinese Han population.
authors.affiliation: Laboratory Department, Jingzhou First People's Hospital, Jingzhou 434000, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Fan
authors.forename: Lu
authors.initials: L
authors.lastname: Huang
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Zhou
authors.forename: Xiaoqian
authors.initials: X
authors.lastname: Zeng
authors.forename: Guining
authors.initials: G
authors.lastname: Li
authors.forename: Pankaj
authors.initials: P
authors.lastname: Deo
authors.forename: Lihua
authors.initials: L
authors.lastname: Hu
authors.forename: Yirong
authors.initials: Y
authors.lastname: Li
mesh_terms: D000328:Adult; D000368:Aged; D002681:China; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005838:Genotype; D015796:HLA-B27 Antigen / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013167:Spondylitis, Ankylosing / Q000235:genetics; D013997:Time Factors
chemical_list: D017931:DNA Primers; D015796:HLA-B27 Antigen