title: Development of multiplex pyrosequencing for HLA-B*57:01 screening using single nucleotide polymorphism haplotype.
abstract: OBJECTIVE
  Abacavir (ABC) is a commonly used nucleoside reverse-transcriptase inhibitor with potent antiviral activity against HIV-1. The US Food and Drug Administration and international HIV treatment guidelines recommend HLA-B*57:01 screening before initiating treatment with ABC. The current standard method for HLA-B*57:01 screening is limited by its high-cost, time-consuming and labour-intensive procedure with the requirement of a specialized laboratory. Our study aims to develop a more reliable screening test by selecting rs3093726 as an additional single nucleotide polymorphism (SNP) to combine with rs2395029 for multiplex pyrosequencing development. It offers high-accuracy, cost-effective and rapid detection.
  
  
  METHODS
  Multiplex pyrosequencing was developed for HLA-B*57:01 screening using rs2395029 and rs3093726 as a surrogate marker and tested in 130 Thai subjects in parallel with singleplex pyrosequencing of each SNP and the standard sequence-based method.
  
  
  CONCLUSIONS
  Multiplex pyrosequencing showed 100% concordance when compared with both singleplex pyrosequencing and standard sequence-based method. This method showed 100% of negative predictive value (NPV), positive predictive value (PPV), specificity and sensitivity.
  
  
  CONCLUSIONS
  Multiplex pyrosequencing is a powerful tool for HLA-B*57:01 screening using the rs2395029 and rs3093726 haplotype genotyping as surrogate marker for this HLA-B. The assay provides accurate, cost-effective and rapid detection of this haplotype. It can be applied for ABC hypersensitivity screening of the Thai population before initiating treatment with ABC.
authors.affiliation: Department of Microbiology and Research, Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
authors.forename: N
authors.initials: N
authors.lastname: Sankuntaw
authors.forename: S
authors.initials: S
authors.lastname: Chantarangsu
authors.forename: W
authors.initials: W
authors.lastname: Chantratita
authors.forename: S
authors.initials: S
authors.lastname: Sungkanuparph
authors.forename: S
authors.initials: S
authors.lastname: Kiertiburanakul
authors.forename: V
authors.initials: V
authors.lastname: Lulitanond
mesh_terms: D019380:Anti-HIV Agents / Q000009:adverse effects* / Q000097:blood; D044466:Asian People / Q000235:genetics; D017931:DNA Primers; D015224:Dideoxynucleosides / Q000009:adverse effects* / Q000097:blood; D004342:Drug Hypersensitivity / Q000235:genetics*; D015235:HLA-B Antigens / Q000235:genetics*; D006239:Haplotypes; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide; D011237:Predictive Value of Tests; D013785:Thailand; D000099227:Abacavir
chemical_list: D019380:Anti-HIV Agents; D017931:DNA Primers; D015224:Dideoxynucleosides; D015235:HLA-B Antigens; D000099227:Abacavir; C484830:HLA-B*57:01 antigen
keywords: adverse effect; genotyping; pharmacogenetics