冷PCR
基因分型
聚合酶链反应
克拉斯
分子生物学
生物
肺癌
突变
突变体
实时聚合酶链反应
基因
基因型
遗传学
医学
点突变
病理
作者
Junjie Zhu,Yanfeng Zhao,Ming Liu,Diego González-Rivas,Xin Xu,Weiping Cai,Hao Qi,Lei Dai,Zijian Wang,Xiao Song,Gening Jiang,Yang Yang
出处
期刊:Small
[Wiley]
日期:2019-01-24
卷期号:15 (9)
被引量:6
标识
DOI:10.1002/smll.201805285
摘要
An accurate genotyping analysis is one of the critical prerequisites for lung cancer targeted therapy. Here, a quantitative polymerase chain reaction (qPCR)-based mutation detection system, mutation-selected amplification-specific system PCR (MASS-PCR), is developed. The specific primers and probes used in MASS-PCR exactly match with the mutant sequence that only allows mutant gene to emit the fluorescence peak. To determine the sensitivity of MASS-PCR, 717 lung cancer specimens, 61 formalin-fixed paraffin-embedded (FFPE) tissues, and 656 fresh reaction tissues are collected and undergo mutation detection of lung cancer driver genes (EGFR, KRAS, BRAF, HER2, MET, ALK, and ROS1). These samples are divided into two groups. Mutations in Group I, which has 631 fresh reaction tissues, are analyzed by MASS-PCR and the amplification refractory mutation system PCR (ARMS-PCR). While group II samples, 25 fresh reaction tissues and 61 FFPE tissues, are screened through MASS-PCR and next-generation sequencing (NGS). All results are verified by direct sequencing. MASS-PCR shows high consistency with ARMS-PCR (kappa value > 0.733) and NGS (kappa value = 0.79) (P < 0.001). For the samples with inconsistent MASS-PCR and ARMS-PCR results, DS results more likely support the MASS-PCR results. These data suggest that MASS-PCR is a convenient, accurate, and economical method for the detection of lung cancer driver gene mutations in clinical practice.
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