中国现代医学杂志2017，Vol.27Issue(22)：13-19,7.DOI:10.3969/j.issn.1005-8982.2017.22.003
建立基于脱氧次黄嘌呤核苷修饰等位基因PCR方法检测EGFR基因热点突变
Establishment of a method for detection of EGFR gene hotspot mutations based on deoxyinosine-modified allele-specific PCR
摘要
Abstract
Objective To establish a method to test the mutations of EGFR gene hotspots L858R and 19 del (2235-2249,2236-2250) based on deoxyinosine-modified allele-specific PCR (dI-AS-PCR).Methods A dI-AS-PCR primer system was designed to detect the mutations of EGFR gene hotspots (L858R and 19 del),which included specific primers modified by deoxyinosine at the n-1 or n-2 position of the primer 3'-terminal,fluorescent probe and internal control primers.Standard substance and quality control samples were built and analyzed by the dI-AS-PCR system.Results The detection limit of dI-AS-PCR was less than 0.1％ under the background of wild type template.The dI-AS-PCR system was used to screen 50 lung cancer samples,in which 9 samples (18％) had L858R mutation of EGFR gene and 14 cases (28％) had 19 de1 mutation.The hotspot mutation rate of EGFR gene was 46％,which was consistent with the results of DNA sequencing (P ＞ 0.05).Conclusions The dI-AS-PCR is a sensitive and specific assay,which could be widely used to detect EGFR mutations,guide patient-specific treatment and monitor the drug resistance of TKI-therapy in clinic.关键词
EGFR热点突变/等位基因PCR/脱氧次黄嘌呤/肺癌Key words
EGFR gene hotspot mutation/allele specific PCR/deoxyinosine/lung cancer分类
医药卫生引用本文复制引用
杨永辉,李辉,朱桂云,李秀武,陈宁,任雪飞..建立基于脱氧次黄嘌呤核苷修饰等位基因PCR方法检测EGFR基因热点突变[J].中国现代医学杂志,2017,27(22):13-19,7.基金项目
河北省卫生和计划生育委员会重点跟踪项目(No:GL2014061) （No:GL2014061）
