等位基因特异扩增法研究中国人CYP2D6中速代谢的相关基因

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57%；29名中速代谢受试者(IMs)以突变型纯合子(m/m)为主占69%；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，可用于CYP2D6中速代谢的检测与研究。

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

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.