title: [Development of the high resolution melting method for genotyping CYP2C9*3 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231)].
abstract: OBJECTIVE
  To establish the high-resolution melting curve (HRM) approach for genotyping CYP2C9*3 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231) and explore its value on estimation of the Warfarin initial dose in comparison with various traditional genotyping methods.
  
  
  METHODS
  CYP2C9*3 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231) genotyping was detected in 100 patients receiving Warfarin therapy by the newly developed HRM method and traditional genotyping methods including PCR-restriction fragment length polymorphism (PCR-RFLP), TaqMan probe and DNA sequencing.
  
  
  RESULTS
  The results of the genotypes obtained from above mentioned methods to detect CYP2C9*3 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231) were similar and consistent. The HRM method is simpler, more economical, and faster compared to the traditional methods. The frequencies of the VKORC1-1639 AA, AG, GG genotypes from these 100 clinical samples were 73 (73%), 23 (23%), 4 (4%), respectively; Frequencies of the CYP2C9 1075 AA, AC, CC genotypes were 94(94%), 6 cases (6%), 0, respectively.
  
  
  CONCLUSIONS
  HRM approach can effectively detect CYP2C9*3 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231) polymorphisms and this method is simpler, more economical, and faster compared to the traditional methods for detecting CYP2C9*3 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231) polymorphisms.
authors.affiliation: Department of Cardiology, Institute of Hypertension, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Guang-lin
authors.initials: GL
authors.lastname: Cui
authors.forename: Hu
authors.initials: H
authors.lastname: Ding
authors.forename: Yu-jun
authors.initials: YJ
authors.lastname: Xu
authors.forename: Chen
authors.initials: C
authors.lastname: Chen
authors.forename: Dao-wen
authors.initials: DW
authors.lastname: Wang
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics*; D065729:Cytochrome P-450 CYP2C9; D004305:Dose-Response Relationship, Drug; D005260:Female; D005615:Freezing; D005838:Genotype; D060005:Genotyping Techniques; D006801:Humans; D008297:Male; D008875:Middle Aged; D006899:Mixed Function Oxygenases / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length; D064417:Vitamin K Epoxide Reductases; D014859:Warfarin / Q000008:administration & dosage* / Q000627:therapeutic use*; D055815:Young Adult
chemical_list: D014859:Warfarin; D006899:Mixed Function Oxygenases; C450260:CYP2C9 protein, human; D065729:Cytochrome P-450 CYP2C9; D001189:Aryl Hydrocarbon Hydroxylases; C578523:VKORC1 protein, human; D064417:Vitamin K Epoxide Reductases