中国医药科学2025，Vol.15Issue(5)：146-149,167,5.DOI:10.20116/j.issn2095-0616.2025.05.34
实时荧光PCR检测非小细胞肺癌EGFR基因突变方法的研究
Research on the detection of EGFR gene mutation in non-small cell lung cancer by real-time fluorescence PCR
摘要
Abstract
Objective To construct a detection method of epidermal growth factor receptor(EGFR)gene mutation based on multiplex PCR,specific primers and TaqMan fluorescence probe technology,and to evaluate its performance indices and clinical application.Methods Primer probes were designed for 21 mutation sites using EGFR gene sequence,and the extraction parameters,reaction system and amplification procedure were optimized.Using EGFR pseudovirus particles and clinical samples,the performance indices and clinical detection of the constructed reagents were evaluated and analyzed.Results The lowest detection limit of EGFR gene mutation detection method constructed this time was 10 ng/reaction system 1.0%of mutant genes could be detected,and the coefficient of variation(CV)of precision ∆Ct was all≤5.0%.In the clinical verification study,the developed reagent and the contrast reagent simultaneously detected 1087 cases of lung cancer samples,the coincidence rate of positive and negative and the total coincidence rate were 100%,and the Kappa coefficient was≥0.75.Conclusion The self-built detection method in this study can achieve high sensitivity detection,and it is equivalent to similar products on the market,meeting industry standards and market demand.关键词
表皮生长因子受体/基因突变/荧光PCR/性能评价/临床验证Key words
Epithelial growth factor receptor/Gene mutation/Fluorescence PCR/Performance evaluation/Clinical verification分类
临床医学引用本文复制引用
杨晓娟,郭志武,马红丽,杨友锋,王浩,王奕江,李振勇..实时荧光PCR检测非小细胞肺癌EGFR基因突变方法的研究[J].中国医药科学,2025,15(5):146-149,167,5.基金项目
江苏省战略性新兴产业发展专项资金(苏发改高技发[2021]77号,苏财建[2021]2号). （苏发改高技发[2021]77号,苏财建[2021]2号）
