Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos, Brazil.
Barretos School of Medicine Dr. Paulo Prata - FACISB, Barretos, Brazil.
Methods Mol Biol. 2021;2279:109-126. doi: 10.1007/978-1-0716-1278-1_9.
Driver mutations in non-small cell lung cancer (NSCLC) have a relevant significance for clinical management. EGFR mutations are the most important predictive biomarkers for NSCLC, although KRAS and BRAF mutations can also be prognostic and predictive biomarkers, respectively. PCR-based approaches followed by sequencing are useful for EGFR, KRAS, and BRAF mutational analysis. Herein, all steps for a PCR-based technique, from DNA isolation from tumor tissue sections to DNA sequencing for genetic analysis of EGFR, KRAS, and BRAF hotspot regions are described.
非小细胞肺癌 (NSCLC) 中的驱动基因突变对临床管理具有重要意义。EGFR 突变是 NSCLC 最重要的预测生物标志物,尽管 KRAS 和 BRAF 突变也分别是预后和预测生物标志物。基于 PCR 的方法,随后进行测序,可用于 EGFR、KRAS 和 BRAF 突变分析。本文描述了一种基于 PCR 的技术的所有步骤,从肿瘤组织切片的 DNA 分离到 EGFR、KRAS 和 BRAF 热点区域的遗传分析的 DNA 测序。