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目的:CYP2C19ml是引起CYP2C19酶活性缺陷的主要等位基因,有83％左右的慢代谢者含有CYP2C19ml等位基因,本文试图建立一步PCR测定CYP2C19ml等位基因的方法。方法:根据等位基因特异扩增(ASA)原理设计两对分别特异扩增野生型等位基因和突变型等位基因的引物,建立了一步PCR测定CYP2C19ml等位基因的方法。结果:对39位随机受试者进行了基因分型研究,发现3位CYP2C19ml纯合子、18位CYP2C19ml杂合子,其余18位为野生型纯合子。结论:说明效法能够用于测定CYP2C19ml等住基因,并且证明该法具有简便、快速和污染少的优点。
OBJECTIVE: CYP2C19ml is the major allele that causes CYP2C19 deficiency. About 83% of CYP2C19ml alleles contain CYP2C19ml allele. This paper attempts to establish a one-step PCR method for CYP2C19ml allele. Methods: According to the principle of allele specific amplification (ASA), two pairs of primers were designed to amplify alleles of wild type and mutant alleles respectively. A method of one - step PCR for CYP2C19ml allele was established. RESULTS: Genotyping of 39 randomized subjects revealed 3 CYP2C19ml homozygotes and 18 CYP2C19ml heterozygotes, and the remaining 18 were wild-type homozygotes. CONCLUSIONS: It is demonstrated that the assay can be used to determine the CYP2C19 ml isozyme, and the method has the advantage of being simple, rapid and less polluting.