Tamakoshi Akiko, Hamajima Nobuyuki, Kawase Haruya, Wakai Kenji, Katsuda Nobuyuki, Saito Toshiko, Ito Hidemi, Hirose Kaoru, Takezaki Toshiro, Tajima Kazuo
Department of Preventive Medicine/Biostatistics and Medical Decision Making, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.
Alcohol Alcohol. 2003 Sep-Oct;38(5):407-10. doi: 10.1093/alcalc/agg096.
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.
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.
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.
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.
采用双引物聚合酶链反应（PCR）结合两对引物（PCR-CTPP）对乙醇脱氢酶β亚基（ADH2）的Arg47His和乙醛脱氢酶2（ALDH2）的Glu487Lys进行基因分型，该方法可通过包含8条引物的单管PCR进行DNA扩增及后续电泳。
测试了多种PCR条件以确定区分这两种多态性的等位基因特异性条带的最佳条件。在最佳PCR条件下，对454名日本健康体检者进行基因分型。
在双管PCR-CTPP的最佳条件下成功扩增出等位基因特异性条带。基因型分布符合Hardy-Weinberg平衡。双管PCR-CTPP基因分型产生的条带比仅针对ADH2进行的PCR-CTPP产生的条带更清晰。
通过这种新开发的双管PCR-CTPP成功对ADH2的Arg47His和ALDH2的Glu487Lys进行了基因分型，这是一种廉价且省时的基因分型工具，将有助于酒精中毒的流行病学研究以及酒精相关疾病的风险评估。
