title: Combination of multiplex PCR and DHPLC-based strategy for CYP2D6 genotyping scheme in Thais.
abstract: OBJECTIVE
  To develop CYP2D6 genotyping scheme for accurate allele calling and reliable estimation of functional allele dosage in Thais.
  
  
  METHODS
  We analyzed CYP2D6 copy numbers by pentaplex PCR coupled with semi-quantitative denaturing high performance liquid chromatography (DHPLC)-based technique. Ten common SNPs were genotyped from CYP2D6 gene product using single base extension (SBE) followed by DHPLC analysis. This detection scheme was compared with real-time PCR and conventional PCR-RFLP for cost-effectiveness.
  
  
  RESULTS
  The distribution of CYP2D6 gene copy numbers in our population ranged from zero (0.69%), one (7.99%), two (60.07%), three (28.13%) and four (3.13%). The most commonly detected SNPs were related to CYP2D6*10 haplotype. CYP2D6*36 in tandem with CYP2D6*10B is the major rearrangement type in Thais (18.75%).
  
  
  CONCLUSIONS
  Multiplex PCR coupled with DHPLC-based strategy is convenient and reliable method for CYP2D6 genotyping offering sufficient allele coverage for Asians. Both cost and analytical time saving were shown and the method could potentially be modified to accommodate CYP2D6 genotyping in other ethnics.
authors.affiliation: Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand.
authors.forename: Payiarat
authors.initials: P
authors.lastname: Suwannasri
authors.affiliation: Division of Molecular Genetics, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkoknoi, Bangkok 10700, Thailand.
authors.forename: Wanna
authors.initials: W
authors.lastname: Thongnoppakhun
authors.affiliation: Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand. Electronic address: ppornpen@chula.ac.th.
authors.forename: Pornpen
authors.initials: P
authors.lastname: Pramyothin
authors.affiliation: Biostatistics and Informatics Laboratory, Genome Institute, National Center for Genetic Engineering and Biotechnology (BIOTEC), Klong Luang, Pathumthani 12120, Thailand.
authors.forename: Anunchai
authors.initials: A
authors.lastname: Assawamakin
authors.affiliation: Division of Molecular Genetics, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkoknoi, Bangkok 10700, Thailand. Electronic address: siclw@mahidol.ac.th.
authors.forename: Chanin
authors.initials: C
authors.lastname: Limwongse
mesh_terms: D044466:Asian People / Q000235:genetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003362:Cost-Benefit Analysis; D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics*; D018628:Gene Dosage; D005838:Genotype; D006801:Humans; D008722:Methods; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D013785:Thailand
chemical_list: D019389:Cytochrome P-450 CYP2D6