Department of Clinical Medicine, Aarhus University, 8000 Aarhus C, Denmark; Department of Pathology, Aarhus University Hospital, 8200 Aarhus N, Denmark.
Department of Clinical Medicine, Aarhus University, 8000 Aarhus C, Denmark; Department of Pathology, Aarhus University Hospital, 8200 Aarhus N, Denmark.
Hum Pathol. 2021 Sep;115:67-75. doi: 10.1016/j.humpath.2021.06.001. Epub 2021 Jun 18.
The aim of the study was to investigate the advantage of implementing next-generation sequencing (NGS) compared with quantitative polymerase chain reaction (qPCR) when performing routine molecular diagnostics in adenocarcinomas of the lung.
The study is a retrospective cross-sectional observational study of 1839 cytological and histological adenocarcinoma biopsies investigated for gene mutations from 2016 to 2018 at the Department of Pathology at Aarhus University Hospital. A total of 1169 samples were analyzed by qPCR for the presence of EGFR hotspot mutations from 2016 to 2017. A total of 670 samples were analyzed with NGS for the presence of EGFR mutations and other gene mutations in 2018.
The average frequency of EGFR mutations in the study population was 11.5%, with the highest frequency found in 2018, where NGS was implemented (10.8% in 2016, 11.5% in 2017, and 12.2% in 2018). Possible therapy resistance markers such as EGFR exon 20 mutations were found more commonly after NGS implementation, the difference being statistically significant (P = .015). In addition, NGS (2018) showed that 40.6% of the samples had KRAS mutations and 6.0% had BRAF mutations, mutations not commonly investigated in lung adenocarcinomas when qPCR is the method of choice. Among the EGFR-mutated samples analyzed with NGS, 13 contained a concurrent EGFR mutation, whereas three and two contained a concurrent KRAS and BRAF mutations, respectively.
With the implementation in a clinical setting, NGS identifies more uncommon but potentially clinically important EGFR mutations, unique combinations of EGFR mutations, and concurrent mutations in KRAS and BRAF.
本研究旨在探讨在肺癌腺癌患者的常规分子诊断中，应用下一代测序（NGS）相较于定量聚合酶链反应（qPCR）的优势。
这是一项回顾性的横断面观察性研究，纳入了 2016 年至 2018 年在奥胡斯大学医院病理科进行基因检测的 1839 例细胞学和组织学腺癌活检。2016 年至 2017 年，1169 例样本通过 qPCR 分析 EGFR 热点突变的存在情况。2018 年，670 例样本通过 NGS 分析 EGFR 突变和其他基因的突变情况。
研究人群中 EGFR 突变的平均频率为 11.5%，其中 2018 年（实施 NGS 时）的频率最高（2016 年为 10.8%，2017 年为 11.5%，2018 年为 12.2%）。实施 NGS 后，更常见到可能导致治疗耐药的标志物，如 EGFR 外显子 20 突变，差异具有统计学意义（P=0.015）。此外，NGS（2018 年）显示 40.6%的样本存在 KRAS 突变，6.0%的样本存在 BRAF 突变，这些突变在选择 qPCR 作为方法时通常不会进行检测。在通过 NGS 分析的 EGFR 突变样本中，13 个样本同时存在 EGFR 突变，3 个样本和 2 个样本分别同时存在 KRAS 和 BRAF 突变。
在临床实施中，NGS 可识别更罕见但具有潜在临床意义的 EGFR 突变、EGFR 突变的独特组合以及 KRAS 和 BRAF 的同时突变。
