Comparison of Targeted Next-Generation Sequencing (NGS) and Real-Time PCR in the Detection of EGFR, KRAS, and BRAF Mutations on Formalin-Fixed, Paraffin-Embedded Tumor Material of Non-Small Cell Lung Carcinoma-Superiority of NGS
Comparison of Targeted Next-Generation Sequencing (NGS) and Real-Time PCR in the Detection of EGFR, KRAS, and BRAF Mutations on Formalin-Fixed, Paraffin-Embedded Tumor Material of Non-Small Cell Lung Carcinoma-Superiority of NGS
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DOI：
10.1002/gcc.22047
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发表时间：
2013-05-01
影响因子：
3.7
通讯作者：
Knuutila, Sakari
中科院分区：
文献类型：
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作者：
Tuononen, Katja;Maki-Nevala, Satu;Knuutila, Sakari
酪氨酸激酶抑制剂治疗方法的发展使得检测癌症患者是否存在影响治疗益处的具有临床意义的基因突变变得非常重要。靶向下一代测序（NGS）通过在单个测试中同时检测各种基因中的多个突变，为诊断目的提供了一种有前途的方法。本研究的目的是通过靶向NGS和常用的实时聚合酶链反应（PCR）方法筛选EGFR、KRAS和BRAF突变，以评估靶向NGS检测突变的可行性。此外，我们旨在通过靶向NGS鉴定潜在的新突变。我们分析了81例非小细胞肺癌患者的福尔马林固定、石蜡包埋（FFPE）肿瘤组织标本。我们观察到靶向NGS和实时PCR之间EGFR、KRAS和BRAF突变检测结果的显著一致性（96.3%至100%）。此外，有针对性的NGS揭示了7个非同义的单核苷酸变异和一个插入-缺失的变化，EGFR不能检测的实时PCR方法。这些变异的潜在临床意义需要在未来的研究中阐明。我们的研究结果支持使用靶向NGS筛选FFPE组织材料中的EGFR、KRAS和BRAF突变。(c)2012 Wiley Periodicals，Inc.
The development of tyrosine kinase inhibitor treatments has made it important to test cancer patients for clinically significant gene mutations that influence the benefit of treatment. Targeted next-generation sequencing (NGS) provides a promising method for diagnostic purposes by enabling the simultaneous detection of multiple mutations in various genes in a single test. The aim of our study was to screen EGFR, KRAS, and BRAF mutations by targeted NGS and commonly used real-time polymerase chain reaction (PCR) methods to evaluate the feasibility of targeted NGS for the detection of the mutations. Furthermore, we aimed to identify potential novel mutations by targeted NGS. We analyzed formalin-fixed, paraffin-embedded (FFPE) tumor tissue specimens from 81 non-small cell lung carcinoma patients. We observed a significant concordance (from 96.3 to 100%) of the EGFR, KRAS, and BRAF mutation detection results between targeted NGS and real-time PCR. Moreover, targeted NGS revealed seven nonsynonymous single-nucleotide variations and one insertion-deletion variation in EGFR not detectable by the real-time PCR methods. The potential clinical significance of these variants requires elucidation in future studies. Our results support the use of targeted NGS in the screening of EGFR, KRAS, and BRAF mutations in FFPE tissue material. (c) 2012 Wiley Periodicals, Inc.