新诊断的转移性非小细胞肺癌（NSCLC）的标准护理是检测ALK、ROS1和表皮生长因子受体（EGFR）三个基因的驱动突变，同时进行免疫组化检测以评估程序性死亡配体-1的表达水平。下一代测序（NGS）越来越多地被应用于标准临床实践中，用于识别可能存在可行动突变的患者，其中可以包括RNA融合测试。目前，NGS突变分层基于欧洲医学肿瘤学会分子靶点临床可操作性分层（ESCAT）I-V和X。我们的目的是分析NSCLC 肿瘤样本中I-V级突变的患病率，为具有转移性疾病的患者提供当前和新的治疗指导。我们使用Oncomine Precision试剂盒（未进行RNA融合测试）进行了NGS检测，该试剂盒检测45个基因中的热点变异体，对2021年6月至2022年3月期间悉尼的六家医院共收集到的210份NSCLC组织样本进行筛查。在我们的队列中，161份（77%）的样本至少发现了一种基因突变，41份（20%）样本同时存在两种或更多突变。其中，I级突变包括210份样本中的42例（20%）EGFR突变（EIA）和5例（3%）MET外显子14剪接突变（EIB）。非I级变异体包括210份样本中的22例（11%）KRAS G12C热点突变（EIIB），另有47例（22%）存在非G12C KRAS（EX）突变（EIIB）。NGS检测显示，在NSCLC中，15%的病例存在II级ESCAT突变。此外，还有46%的患者表现出潜在的III级和IV级突变，目前这些突变正在早期临床试验中进行研究。除了识别适合临床证实的标准治疗方案的基因变异的患者，45基因NGS面板在识别可能可行但非I级突变方面具有重要潜力，这些突变可能会成为NSCLC的未来标准临床实践。© 2023 Royal Australasian College of Physicians.

The standard of care in newly diagnosed metastatic non-small cell lung cancer (NSCLC) is to test for aberrations in three genes for driver mutations - ALK, ROS1 and epidermal growth factor receptor (EGFR) - and also for immunohistochemistry to be performed for programmed death-ligand 1 expression level. Next-generation sequencing (NGS), with or without RNA fusion testing, is increasingly used in standard clinical practice to identify patients with potentially actionable mutations. Stratification of NGS mutation tiers is currently based on the European Society of Medical Oncology Scale for Clinical Actionability of Molecular Targets (ESCAT) Tiers I-V and X.Our aim was to analyse NSCLC tumour samples for the prevalence of Tiers I-V mutations to establish guidance for current and novel treatments in patients with metastatic disease.NGS was performed employing the Oncomine Precision Assay (without RNA fusion testing) that interrogates DNA hotspot variants across 45 genes to screen 210 NSCLC tissue samples obtained across six Sydney hospitals between June 2021 and March 2022.In our cohort, 161 of 210 (77%) had at least one gene mutation identified, with 41 of 210 (20%) having two or more concurrent mutations. Tier I mutations included 42 of 210 (20%) EGFR mutations (EIA) and five of 210 (3%) MET exon 14 skipping mutations (EIB). Non-Tier I variants included 22 of 210 (11%) KRAS G12C hotspot mutations (EIIB), with a further 47 of 210 (22%) having non-G12C KRAS (EX) mutations. NGS testing revealed an additional 15% of cases with Tier II ESCAT mutations in NSCLC. Forty-six percent of patients also demonstrated potential Tier III and IV mutations that are currently under investigation in early-phase clinical trials.In addition to identifying patients with genomic alterations suitable for clinically proven standard-of-care therapeutic options, the 45-gene NGS panel has significant potential in identifying potentially actionable non-Tier 1 mutations that may become future standard clinical practice in NSCLC.© 2023 Royal Australasian College of Physicians.