Stüven T, Griese E U, Kroemer H K, Eichelbaum M, Zanger U M
Dr Margarete Fischer-Bosch-Institut für Klinische Pharmakologie, Stuttgart, Germany.
Pharmacogenetics. 1996 Oct;6(5):417-21. doi: 10.1097/00008571-199610000-00005.
The CYP2D6 gene on human chromosome 22 encodes a cytochrome P450 responsible for oxidative metabolism of over 30 clinically used drugs. The CYP2D6 gene is highly polymorphic with more than 20 alleles described to date. Some of these harbour loss-of-function mutations which lead to the poor metabolizer phenotype in 5-10% of Caucasians. These individuals are at increased risk of suffering from adverse side effects or to experience therapeutic failure following drug treatment. Phenotype determination requires ingestion of a probe drug and has other inherent problems. Due to the increasing number of alleles known, comprehensive CYP2D6 genotyping using the conventional assays has become cumbersome and time consuming. We have therefore developed a streamlined and more rapid CYP2D6 genotyping procedure. Use of long distance PCR allowed the amplification of a 4666 bp fragment which contains the entire CYP2D6 gene. The 4.7 kb fragment serves as a template for a multiplex allele-specific PCR assay to simultaneously identify the five PM-associated alleles, CYP2D6*3 (A), *4 (B), *6 (T), *7 (E), and 8 (G). Together with the CYP2D6 deletion allele CYP2D65 (D), which can be detected in a separate PCR assay, these alleles are responsible for the PM phenotype in approximately 99% of Caucasian individuals. We tested the reliability of the procedure by analysing DNA from more than 80 individuals with known CYP2D6 genotypes. Twelve different genotypes were present among these samples and all of them were correctly identified.
人类22号染色体上的CYP2D6基因编码一种细胞色素P450，负责30多种临床常用药物的氧化代谢。CYP2D6基因具有高度多态性，迄今已描述了20多个等位基因。其中一些携带功能丧失突变，导致5-10%的高加索人出现代谢不良者表型。这些个体在药物治疗后出现不良反应或治疗失败的风险增加。表型测定需要摄入探针药物，并且存在其他固有问题。由于已知等位基因数量的增加，使用传统检测方法进行全面的CYP2D6基因分型变得繁琐且耗时。因此，我们开发了一种简化且更快速的CYP2D6基因分型程序。使用长距离PCR可以扩增出一个包含整个CYP2D6基因的4666 bp片段。这个4.7 kb的片段作为多重等位基因特异性PCR检测的模板，用于同时鉴定五个与代谢不良相关的等位基因，即CYP2D6*3（A）、*4（B）、6（T）、7（E）和8（G）。连同可以在单独的PCR检测中检测到的CYP2D6缺失等位基因CYP2D65（D），这些等位基因在大约99%的高加索个体中导致代谢不良表型。我们通过分析来自80多名已知CYP2D6基因型个体的DNA来测试该程序的可靠性。这些样本中存在12种不同的基因型，并且所有基因型都被正确鉴定。
