title: Direct genotyping of Cytochrome P450 2A6 whole gene deletion from human blood samples by the SmartAmp method.
abstract: BACKGROUND
  Genetic polymorphisms of the human CYP2A6 gene are considered to be a determinant of smoking behavior and tobacco-related lung cancer risk in male Japanese smokers. We developed a SmartAmp-based genotyping method to detect whole deletion of the CYP2A6 gene directly from blood samples without DNA isolation.
  
  
  METHODS
  We validated the new method using CYP2A plasmids, 48 genomic DNA samples and 25 blood samples by utilizing the SmartAmp method, a unique isothermal DNA amplification process.
  
  
  RESULTS
  This method could discriminate the CYP2A6 gene from highly homologous CYP2A7 and CYP2A13 genes. CYP2A6*1 (wild-type) and CYP2A6*4 (whole gene deletion) were determined by the new method in perfect accordance with sequence analysis data.
  
  
  CONCLUSIONS
  A SmartAmp assay for genotyping the CYP2A6 gene was developed, and the reliability of the method was validated using the conventional PCR method.
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan.
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Azuma
authors.forename: Alexander
authors.initials: A
authors.lastname: Lezhava
authors.forename: Makiko
authors.initials: M
authors.lastname: Shimizu
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Kimura
authors.forename: Yuri
authors.initials: Y
authors.lastname: Ishizu
authors.forename: Toshihisa
authors.initials: T
authors.lastname: Ishikawa
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Kamataki
authors.forename: Yoshihide
authors.initials: Y
authors.lastname: Hayashizaki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yamazaki
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000097:blood* / Q000172:deficiency / Q000235:genetics*; D065637:Cytochrome P-450 CYP2A6; D004247:DNA / Q000097:blood / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D017353:Gene Deletion; D005838:Genotype; D006358:Hot Temperature; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D013997:Time Factors
chemical_list: D017931:DNA Primers; D004247:DNA; D001189:Aryl Hydrocarbon Hydroxylases; C585710:CYP2A6 protein, human; D065637:Cytochrome P-450 CYP2A6