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- 2017
Real-time PCR、焦磷酸测序及基因芯片快速检测ALDH2*2基因多态性
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Abstract:
摘要：目的 建立ALDH2基因多态性的快速检测方法，研究ALDH2基因多态性是否与中国人群饮酒相关。方法 Real-time PCR、焦磷酸测序仪及基因芯片用于检测ALDH2*2多态性，302个志愿者参与这项研究，评估饮酒行为与ALDH2*2多态性之间的关系。结果 Real-time PCR、焦磷酸测序及基因芯片能成功的鉴别ALDH2*2多态性位点，ALDH2*2等位基因的频率在中国人群中为0.2036，其中在饮酒人群中占0.1633，而在非饮酒人群中为0.2783(P=0.001)，ALDH2*1/*2与ALDH2*2/*2基因型频率在非饮酒人群中为0.4528，而在饮酒人群中占0.3265(P=0.030)，中国人群ALDH2*2等位基因的频率要远高于欧洲及美洲，并与其他亚洲人群存在显著性差异。结论 建立的Real-time PCR、焦磷酸测序及基因芯片技术方法快速、精准、高通量、方便，适合用于检测ALDH2*2基因多态性位点，ALDH2*2等位基因能防止酗酒，中国人群ALDH2*2等位基因的频率与其他种族人群显著不同。
ABSTRACT: Objective To establish a rapid molecular method for the detection of aldehyde dehydrogenase-2 (ALDH2) and investigate the gene polymorphisms of ALDH2*2 and determine whether the polymorphic ALDH2 gene is associated with drinking behavior in a Chinese population. Methods The gene polymorphisms of ALDH2*2 were detected using pyrosequencing, TaqMan Real-time PCR and GeneChip microarray technologies; genotyping of 302 volunteers was performed to assess their genetic associations with alcohol use behavior.Results We developed pyrosequencing, TaqMan Real-time PCR and GeneChip microarray methods to identify ALDH2*2 polymorphisms. The allele frequency of ALDH2*2 was 20.36% in the Chinese population: 16.33% in the alcoholic group and 27.83% in non-drinkers (P=0.001). In contrast, the genotype frequency of heterozygous ALDH2*1/*2 plus homozygous ALDH2*2/*2 was 45.28% in non-drinkers and 32.65% in the alcoholics group (P=0.030). Allele frequency of ALDH2 genotypes differed significantly between our Chinese sample and other ethnic groups in Asia, and it was significantly higher than that in European and American countries.Conclusion The developed pyrosequencing, TaqMan Real-time PCR and GeneChip microarray methods are rapid, accurate, high-throughput, convenient, and reliable for detecting ALDH2 polymorphisms. ALDH2？？*2 gene can protect against the development of alcoholism. The allele frequency of ALDH2 in this Chinese population differs from that in other ethnic groups
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