title: Duplex polymerase chain reaction with confronting two-pair primers (PCR-CTPP) for genotyping alcohol dehydrogenase beta subunit (ADH2) and aldehyde dehydrogenase 2 (ALDH2).
abstract: OBJECTIVE
  Alcohol dehydrogenase beta subunit (ADH2) Arg47His and aldehyde dehydrogenase 2 (ALDH2) Glu487Lys were genotyped by a duplex polymerase chain reaction (PCR) with confronting two-pair primers (PCR-CTPP), which allows DNA amplification with one-tube PCR including eight primers, and subsequent electrophoresis.
  
  
  METHODS
  Several PCR conditions were tested to establish the optimal conditions for distinguishing the allele-specific bands for the two polymorphisms. Under the optimal PCR conditions, 454 Japanese health check-up examinees were genotyped.
  
  
  RESULTS
  The allele-specific bands were successfully amplified under the optimal conditions of the duplex PCR-CTPP. The genotype distributions were within the Hardy-Weinberg equilibrium. The bands produced by the duplex PCR-CTPP genotyping were clearer than those produced by PCR-CTPP, conducted solely for ADH2.
  
  
  CONCLUSIONS
  ADH2 Arg47His and ALDH2 Glu487Lys were successfully genotyped by this newly developed duplex PCR-CTPP, an inexpensive and time-saving genotyping tool, which will be useful in epidemiological studies on alcoholism, as well as risk estimation of alcohol-related diseases.
authors.affiliation: Department of Preventive Medicine/Biostatistics and Medical Decision Making, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Tamakoshi
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Hamajima
authors.forename: Haruya
authors.initials: H
authors.lastname: Kawase
authors.forename: Kenji
authors.initials: K
authors.lastname: Wakai
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Katsuda
authors.forename: Toshiko
authors.initials: T
authors.lastname: Saito
authors.forename: Hidemi
authors.initials: H
authors.lastname: Ito
authors.forename: Kaoru
authors.initials: K
authors.lastname: Hirose
authors.forename: Toshiro
authors.initials: T
authors.lastname: Takezaki
authors.forename: Kazuo
authors.initials: K
authors.lastname: Tajima
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000426:Alcohol Dehydrogenase / Q000235:genetics*; D000444:Aldehyde Dehydrogenase / Q000235:genetics*; D000071396:Aldehyde Dehydrogenase, Mitochondrial; D016009:Chi-Square Distribution; D017931:DNA Primers / Q000235:genetics*; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D009692:Nucleic Acid Heteroduplexes; D000426:Alcohol Dehydrogenase; C096127:ALDH2 protein, human; D000444:Aldehyde Dehydrogenase; D000071396:Aldehyde Dehydrogenase, Mitochondrial