Department of Pathology, Haartman Institute, University of Helsinki, Helsinki, Finland.
Genes Chromosomes Cancer. 2013 May;52(5):503-11. doi: 10.1002/gcc.22047. Epub 2013 Jan 30.
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.
酪氨酸激酶抑制剂治疗的发展使得检测影响治疗获益的临床显著基因突变变得非常重要。靶向下一代测序（NGS）通过在单次检测中同时检测多个基因中的多个突变，为诊断目的提供了一种有前途的方法。我们的研究旨在通过靶向 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 名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 之间的 EGFR、KRAS 和 BRAF 突变检测结果具有显著的一致性（从 96.3%到 100%）。此外，靶向 NGS 揭示了 EGFR 中的七个非同义单核苷酸变异和一个实时 PCR 方法无法检测到的插入缺失变异。这些变体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在 FFPE 组织材料中使用靶向 NGS 筛选 EGFR、KRAS 和 BRAF 突变。
