等位基因特异扩增法研究中国人CYP2D6中速代谢的相关基因

ALLELE SPECIFIC AMPLIFICATON FOR CYP2D6 GENE RELATED TO INTERMEDIATE METABOLIZER IN CHINESE SUBJECTS

目的 建立CYP2D6 10B的等位基因特异扩增法 (ASA PCR) ,以探讨中国人CYP2D6中速代谢的基因分型。方法 采用两步扩增法得到CYP2D6 10B等位基因特异片段 ,分析健康中国汉族人CYP2D6 10B等位基因 ,并探讨基因分型结果与右美沙芬表型分型结果的相关性。结果 35名表型为极快代谢受试者 (VEMs)中 ,CYP2D6 10B以杂合子 (wt/m)为主占 5 7% ;2 9名中速代谢受试者 (IMs)以突变型纯合子 (m/m)为主占 6 9% ;慢代谢受试者 (PM)基因型为m/m。CYP2D6 10Bm/m组的MR明显大于wt/m组和野生型组 (wt/wt)。结论 ASA PCR法有快速、准确的优点 。

AIM To establish an allele specific PCR amplification (ASA PCR) for determination of the genotype of CYP2D6 *10B polymorphism in Chinese subjects. METHODS CYP2D6 *10B alleles of 65 healthy Chinese subjects were analyzed by a two step PCR assay and the correlation of genotype and phenotype was studied. RESULTS There were 20 CYP2D6 *10B heterozygous genotypes subjects (wt/m) in 35 very extensive metabolizers (VEMs), which consisted the major part of VEM subjects (57%). Meanwhile, 20 subjects consisting 69% of 29 intermediate metabolizers were CYP2D6 *10B homozygous mutant genotypes ( m/m ). The poor metabolizer was also m/m. The metabolic ratio of CYP2D6 *10B m/m subjects were larger than wt/m and wild type, the values were -1 49 ±0 54, -2 20 ±0 49 and -2 47 ±0 61 ( P <0 01). CONCLUSION PCR ASA was shown to be a rapid and specific method. It can be used to study the genetic polymorphism, especially CYP2D6 intermediate metabolism.