TaqMan Real-Time Polymerase Chain Reaction and Pyrosequencing using Single Nucleotide Polymorphism Protocol for Rapid Determination of ALDH2 *2 in a Chinese Population

Abstract

Purpose: To establish a rapid molecular method for the detection of aldehyde dehydrogenase 2 (ALDH2), and determine whether the polymorphic ALDH2 gene is associated with drinking behavior in a Chinese population.Methods: The gene polymorphism of ALDH2 *2 was detected using pyrosequencing and TaqMan realtime polymerase chain reaction (PCR) techniques. Genotyping of 176 volunteers were performed at The Third Hospital of Wuhan, Hubei, China. Genetic associations with alcohol use behavior was assessed.Results: Pyrosequencing and TaqMan real time PCR methods were successfully developed to identify ALDH2 *2 polymorphisms. The allele frequencies of ALDH2 *2 were 21.31 % in the Chinese population. While the genotype frequency of heterozygous ALDH2 *1/ *2 plus homozygous ALDH2 *2/ *2 was 46.67 % in non-drinkers, it was 30.17 % in the alcoholics group (p = 0.030). The allele frequency of ALDH2 *2 was 15.09 % in the alcoholics group and 28.33 % among non-drinkers (p = 0.011). There were substantial differences in allele frequencies of ALDH2 genotypes between this study’s Chinese sample and other ethnic groups in Asia; the frequency of ALDH2 *2 allele was significantly higher than that in European and Americans.Conclusion: The developed pyrosequencing and TaqMan real time PCR methods are rapid, accurate, high-throughput, convenient, and reliable for detecting ALDH2 polymorphisms. ALDH2 *2 gene protects against the development of alcoholism. The frequencies of ALDH2 in this Chinese population differ from those of other ethnic populations.Keywords: Aldehyde dehydrogenase 2, Polymorphism, TaqMan real time Polymerase Chain Reaction, Pyrosequencing, Allele, Alcoholics, Genotype frequency