Rapid and reliable genotyping of HLA-B*27 in the Chinese Han population using a duplex real-time TaqMan PCR assay

Wen Fan, Lu Huang, Zhiming Zhou, Xiaoqian Zeng, Guining Li, Pankaj Deo, Lihua Hu, Yirong Li

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9 Citations (Scopus)

Abstract

Objectives: To develop a duplex real-time TaqMan PCR assay for genotyping HLA-B*27 in the Chinese Han population. Design and 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.

Original languageEnglish
Pages (from-to)106-111
Number of pages6
JournalClinical Biochemistry
Volume45
Issue number1-2
DOIs
Publication statusPublished - 1 Jan 2012
Externally publishedYes

Keywords

  • 27
  • Ankylosing spondylitis
  • Genotype
  • HLA-B

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