限制性片段长度多态性聚合酶链反应法检测人细胞色素氧化酶2D6药物代谢相关基因位点多态性

Detection of CYP2D6 drug metabolism related locus polymorphism by PCR-RFLP

目的建立测定人细胞色素氧化酶2D6(CYP2D6)基因多态性的限制性片段长度多态性聚合酶链反应(polymerase chain reaction restriction fragment length polymorphism,PCR RFLP)检测方法。方法通过增加PCR扩增体系中DNA模板上样体积、降低引物浓度以及减少限制性内切酶的用量等方法改进、优化实验条件,建立以PCR RFLP法为基础的肝药酶CYP2D6基因型检测方法。结果采用PCR RFLP法能够检测CYP2D6?2、CYP2D6?10和CYP2D6?14基因型,且过程简便,经济实用。结论该方法检测CYP2D6药物代谢相关基因位点基因型具有简便、高效、准确和经济的特点,可在临床及实验室推广使用。

Objective To establish a method for the determination of human cytochrome P4502D6(CYP2D6)gene polymorphism by polymerase chain reaction restriction fragment length polymorphism(PCR RFLP).Methods The experimental conditions were improved and optimized by increasing the sample size of DNA template,reducing the primer concentration and the amount of re striction endonuclease in PCR amplification system.The method for detecting the CYP2D6 genotype of liver drug enzyme based on PCR RFLP method was established.Results The CYP2D6?2,CYP2D6?10 and CYP2D6?14 genotypes could be detected by PCR RFLP method,and the process was simple and economical.Conclusion PCR RFLP method is simple,efficient,accurate and economical for detecting genotypes of CYP2D6 drug related gene loci and can be used in clinical and laboratory applications.