数字聚合酶链反应
T790米
肺癌
表皮生长因子受体
离子半导体测序
吉非替尼
埃罗替尼
生物
DNA测序
分子生物学
突变
癌症研究
聚合酶链反应
癌症
DNA
医学
遗传学
基因
病理
作者
Katsuhiro Masago,Shiro Fujita,Akito Hata,Chiyuki Okuda,Yuko Yoshizumi,Reiko Kaji,Nobuyuki Katakami,Yukio Hirata,Yasushi Yatabe
摘要
The aim of this study was to compare the accuracy of the QuantStudio 3D Digital polymerase chain reaction (dPCR) system and a PCR‐based next generation sequencing (NGS) system for detecting a secondary mutation in the epidermal growth factor receptor (EGFR) gene T790M in non‐small cell lung cancer (NSCLC) patients previously diagnosed with EGFR ‐activating mutations. Twenty‐five patients with NSCLC previously treated with EGFR‐TKIs were examined. The patients were treated daily with either erlotinib or gefitinib. New biopsies, followed by DNA sequencing on an Ion Torrent systems using the Ion Torrent AmpliSeq Cancer Hotspot Panel and dPCR were performed. A comparison of NGS, sensitive PCR, and dPCR revealed that the sensitivities of NGS and dPCR were similar in this study. As well, T790M was detected in as low as about 5% of mutant allelic frequencies, which represented 5% of the total reads on site mapped reads in NGS and greater than 5% of the dPCR reads, which represented mutant and wild type copies. The strategy in which NGS sequencing is followed by revealed acquired mutation with dPCR may be a reasonable one. We demonstrated the utility of combining NGS and dPCR as a tool for monitoring T790M. NGS and dPCR with formalin‐fixed paraffin‐embedded (FFPE) specimens might become a standard genomic test for exploring acquired resistance to targeted molecular medicines.
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