Developing a New qPCR‐Based System for Screening Mutation

• DOI: 10.1002/smll.201805285
• Publication Date: 2019-01-24T23:11:07Z
• Authors: Junjie Zhu; Yanfeng Zhao; Ming Liu; Diego Gonzalez‐Rivas; Xinnan Xu; Weijing Cai; Huiwei Qi; Lei Dai; Zijian Wang; Xiao Song; Gening Jiang; Yang Yang
• Keywords: COLD-PCR

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), developed. The specific primers and probes used in MASS-PCR exactly match with mutant sequence that only allows gene to emit fluorescence peak. To determine sensitivity MASS-PCR, 717 specimens, 61 formalin-fixed paraffin-embedded (FFPE) tissues, 656 fresh tissues are collected undergo driver genes (EGFR, KRAS, BRAF, HER2, MET, ALK, ROS1). These samples divided into two groups. Mutations Group I, which has 631 analyzed by amplification refractory (ARMS-PCR). While group II samples, 25 FFPE screened through next-generation sequencing (NGS). All results verified direct sequencing. shows high consistency ARMS-PCR (kappa value > 0.733) NGS = 0.79) (P < 0.001). For inconsistent results, DS more likely support results. data suggest convenient, accurate, economical method mutations clinical practice.