Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

TaqMan real-time polymerase chain reaction and pyrosequencing using single nucleotide polymorphism protocol for rapid determination of aldh2 *2 in a chinese population

Ju-yi Li¹, Jin-hu Wu², Yan Zhang², Xiu-fang Wang², Jie Jin², Yi Wang¹

¹Department of Pharmacy, The Central Hospital of Wuhan; ²The Third Hospital of Wuhan, Wuhan, Hubei Province, China.

For correspondence:- Yi Wang Email: wxfls727@163.com

Received: 6 March 2015 Accepted: 26 July 2015 Published: 29 September 2015

Citation: Li J, Wu J, Zhang Y, Wang X, Jin J, Wang Y. TaqMan real-time polymerase chain reaction and pyrosequencing using single nucleotide polymorphism protocol for rapid determination of aldh2 *2 in a chinese population. Trop J Pharm Res 2015; 14(9):1679-1684 doi: 10.4314/tjpr.v14i9.19

© 2015 The authors.
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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 real-time 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