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Alcohol and Alcoholism Vol. 38, No. 5, pp. 407-410, 2003
© 2003 Medical Council on Alcohol

DUPLEX POLYMERASE CHAIN REACTION WITH CONFRONTING TWO-PAIR PRIMERS (PCR–CTPP) FOR GENOTYPING ALCOHOL DEHYDROGENASE ß SUBUNIT (ADH2) AND ALDEHYDE DEHYDROGENASE 2 (ALDH2)

Akiko Tamakoshi, Nobuyuki Hamajima*, Haruya Kawase, Kenji Wakai, Nobuyuki Katsuda1, Toshiko Saito2, Hidemi Ito2, Kaoru Hirose2, Toshiro Takezaki2 and Kazuo Tajima2

Department of Preventive Medicine/Biostatistics and Medical Decision Making, Nagoya University Graduate School of Medicine, Nagoya,
1 Nagoya Kita Health Center, Nagoya and
2 Division of Epidemiology and Prevention, Aichi Cancer Center Research Institute, Nagoya, Japan

(Received 28 January 2003; first review notified 17 March 2003; in revised form 3 April 2003; accepted 11 April 2003)

Aims: Alcohol dehydrogenase ß 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.


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