现代检验医学杂志2012，Vol.27Issue(1)：66-70,5.DOI:10.3969/j.issn.1671-7414.2012.01.021
PCR-Pyrosequencing 快速检测药物代谢酶基因CYP2C19多态性方法的建立及应用
Development of a PCR-Pyrosequencing Method for Detecting Rapidly the Genetic Polymorphism of CYP2C19 about Drug Metabolizing Enzymes Genes
摘要
Abstract
Objective To develop a new high throughput method for detecting genetic polymorphism of CYP2C19 about drug metabolism enzyme genes rapidly. Methods 96 examples of genomic DNA from human peripheral blood was extracted, and amplified with biotinylated primer to obtain single-stranded templates for pyrosequencing. Then, the single-stranded templates were subjected to Pyrosequencing analysis using PyroMark ID instrument. Simultaneously,Sanger sequencing was also applied to sequence the products as a control to check the reliability of the pyrosequencing result. Results The results displayed that CYP2C19 related genetic polymorphism could be sucessfuly detected using pyrosequencing method,Repeat test and reliability test indicated that the agreement between the pyrosequencing and the Sanger sequencing methods was 100%. Meanwhile,multiple PCR products combined with pyrosequencing detection through design the pyrosequencing primerscom-pared with a single-tube sites pyrosequencing methods, it was much more economical and timesaving. Conclution All of these demonstrated that pyrosequencing could accurately and rapidly detect the genetic polymorphism of drug metabolism-related genes with high throughput. It has a great value in personalized medical treatment and could be extended to the other genetic diseases.关键词
CYP2C19/焦磷酸测序Key words
CYP2C19/ pyrosequencing分类
生物科学引用本文复制引用
施宏,虞闰六,马金飞,任绪义..PCR-Pyrosequencing 快速检测药物代谢酶基因CYP2C19多态性方法的建立及应用[J].现代检验医学杂志,2012,27(1):66-70,5.
