Department of Pathology, Shanxi Province Cancer Hospital/Shanxi Hospital Affiliated to Cancer Hospital, Chinese Academy of Medical Sciences/Cancer Hospital Affiliated to Shanxi Medical University, Taiyuan, 030013, Shanxi, China.
First Clinical Medical School, Shanxi Medical University, Taiyuan, 030001, Shanxi, China.
J Cancer Res Clin Oncol. 2023 Sep;149(11):8573-8580. doi: 10.1007/s00432-023-04794-z. Epub 2023 Apr 25.
Cell-free circulating tumor DNA (ctDNA) in plasma enables rapid and repeat testing of actionable mutations. Next-generation sequencing (NGS) is an attractive platform for multiplex sequencing capabilities compared to traditional methods such as PCR. The purpose of this study is to evaluate the value of the NGS-based ctDNA assay and to identify the genomic alteration profile of ctDNA in real-world Chinese non-small cell lung (NSCLC) patients.
In total, 294 Chinese patients with pathological diagnosis of Phase III-IV NSCLC were enrolled. 3-4 mL peripheral blood was collected and NGS-based analysis was carried out using a 20-gene panel. The analytical sensitivity and specificity of ctDNA NGS-based assay was validated using droplet digital PCR (ddPCR).
We have tested 570 sites from 286 samples using ddPCR, which included 108 positive sites and 462 negative sites from NGS results, and the concordance rate was 99.8% (418/419) for single-nucleotide variants (SNVs) and 96.7% (146/151) for insertions and deletions (InDels). The most frequent genes were TP53 (32%), EGFR (31.97%), KRAS (6.46%), PIK3CA (4.76%), and MET (4.08%). Exon 19 deletion (19del) was the most common alteration in EGFR and G12C was the most common alteration in KRAS. Furthermore, the detection rate of TP53 was higher in the male and patients with squamous cell carcinoma. We also found the prevalence of TP53 in L858R was higher than in 19del (61.29% vs. 40%; p = 0.1115).
The results indicate that the results of NGS-based ctDNA assay are highly consistent with ddPCR. In Chinese NSCLC patients, TP53 mutation was more frequently associated with male and squamous cell carcinoma. The prevalence of concomitant mutations in L858R may be different from that in 19del.
血浆中的无细胞循环肿瘤 DNA（ctDNA）可实现对可操作突变的快速重复检测。与传统方法（如 PCR）相比，下一代测序（NGS）是一种具有吸引力的多通道测序平台。本研究旨在评估基于 NGS 的 ctDNA 检测的价值，并确定真实世界中中国非小细胞肺癌（NSCLC）患者 ctDNA 的基因组改变谱。
共纳入 294 例经病理诊断为 III-IV 期 NSCLC 的中国患者。采集 3-4ml 外周血，采用 20 基因panel 进行基于 NGS 的分析。使用液滴数字 PCR（ddPCR）验证了 ctDNA NGS 检测的分析灵敏度和特异性。
我们使用 ddPCR 测试了 286 个样本中的 570 个位点，其中包括来自 NGS 结果的 108 个阳性位点和 462 个阴性位点，单核苷酸变异（SNVs）的一致性率为 99.8%（418/419），插入和缺失（InDels）的一致性率为 96.7%（146/151）。最常见的基因是 TP53（32%）、EGFR（31.97%）、KRAS（6.46%）、PIK3CA（4.76%）和 MET（4.08%）。EGFR 中最常见的突变为外显子 19 缺失（19del），KRAS 中最常见的突变为 G12C。此外，TP53 检测率在男性和鳞状细胞癌患者中较高。我们还发现 TP53 在 L858R 中的发生率高于 19del（61.29% vs. 40%；p=0.1115）。
结果表明，基于 NGS 的 ctDNA 检测结果与 ddPCR 高度一致。在中国 NSCLC 患者中，TP53 突变与男性和鳞状细胞癌的相关性更高。L858R 中伴随突变的发生率可能与 19del 不同。
