title: New pharmacogenetic test for detecting an HLA-A*31: 01 allele using the InvaderPlus assay.
abstract: OBJECTIVE
  Carbamazepine (CBZ) is widely used for the treatment of epilepsy and other neurological disorders. However, 3-5% of CBZ-treated individuals suffer from cutaneous adverse drug reactions (cADRs). Recently, in a genome-wide association study, HLA-A*31:01 has been reported to be a strong genetic marker for CBZ-induced cADRs in both Japanese and European populations. As most of the available methods for HLA genotyping are laborious, the development of a simple and rapid genotyping method for HLA-A*31:01 is desirable from the viewpoint of a clinical pharmacogenetic test.
  
  
  METHODS
  More than 1700 sequences for HLA-A alleles were obtained from the MHC database of the National Center for Biotechnology Information (dbMHC). Several HLA-A*31:01-discriminating single-nucleotide polymorphisms were selected. These SNPs were used for sequence-specific primer PCR (SSP-PCR) and for the target site of the Invader reaction. By combining SSP-PCR with a target-specific Invader reaction, we designed two sets of primers/probes for HLA-A*31:01 allele detection. The performance of both sets was evaluated using 90 Asian HapMap samples. Further evaluation was carried out using another 376 Japanese samples and 90 CEU (European) and 90 YRI (African) HapMap samples.
  
  
  RESULTS
  Our assay specifically detected an HLA-A*31:01 allele in a total of 466 individuals of the Asian population. Furthermore, the assay correctly identified HLA-A*31:01-positive carriers from the CEU and the YRI population, respectively, implying that the assay has potential for application to other ethnic groups.
  
  
  CONCLUSIONS
  We developed a new HLA-A*31:01-detecting method by a combination of SSP-PCR with target-specific InvaderPlus technology. As our assay is rapid and accurate, it is hoped that this method will be used in a pharmacogenetic test in a clinical setting to avoid CBZ-induced cADRs.
authors.affiliation: Laboratory for Genotyping Development, Center for Genomic Medicine, RIKEN, Yokohama, Kanagawa, Japan. aoki@src.riken.jp
authors.forename: Masayuki
authors.initials: M
authors.lastname: Aoki
authors.forename: Naoya
authors.initials: N
authors.lastname: Hosono
authors.forename: Sadaaki
authors.initials: S
authors.lastname: Takata
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Kamatani
authors.forename: Michiaki
authors.initials: M
authors.lastname: Kubo
mesh_terms: D000483:Alleles; D044466:Asian People / Q000235:genetics; D017931:DNA Primers; D005006:Ethnicity / Q000235:genetics; D015234:HLA-A Antigens / Q000235:genetics*; D006801:Humans; D010597:Pharmacogenetics / Q000379:methods*; D011933:Reagent Kits, Diagnostic
chemical_list: D017931:DNA Primers; D015234:HLA-A Antigens; C506273:HLA-A*31:01 antigen; D011933:Reagent Kits, Diagnostic