Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Department of Infectious Diseases, Institute for Viral Hepatitis, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, 400010, China.
Clinical Molecular Medicine Testing Center, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
J Cancer Res Clin Oncol. 2022 Feb;148(2):321-330. doi: 10.1007/s00432-021-03818-w. Epub 2021 Oct 24.
By comparing the detection rate and type of targeted gene mutations in non-small cell lung cancer (NSCLC) between amplification refractory mutation system PCR (ARMS-PCR) and next-generation sequencing (NGS), the characteristics and application advantages of non-small cell lung cancer detection are explained, providing a basis for clinicians to effectively select the corresponding detection methods.
The cases of targeted genes for lung cancer were selected from the First Affiliated Hospital of Chongqing Medical University from January 2016 to October 2020. A sample of 4467 cases was selected, and they were diagnosed with NSCLC by Pathological biopsy. Sample sources include surgical resection, bronchoscope biopsy, metastatic biopsy, blood, sputum, cytology of pleural effusion. Among them, 3665 cases were detected by ARMS-PCR technique, and 802 cases were detected by NGS technology. The detection rate and type of ARMS-PCR and NGS techniques for EGFR gene mutations (including exon 18, exon 19, exon 20, exon 21 and so on) in different NSCLC samples were compared, respectively.
The total mutation rate of EGFR gene detected by ARMS-PCR was 47.6% while 42.4% detected by NGS which indicated that there was a significant difference between the two methods in detecting total mutation of EGFR gene (P < 0.001). In different exons, the EGFR mutation rate detected by two methods is various. The mutation rate of exon 19 by ARMS-PCR detection was evidently higher than that of NGS detection, while the mutation rate of exons 20 and 21 by ARMS-PCR detection were statistically significantly lower than that of NGS detection. Moreover, the multiple mutation rate detected by NGS was 16.3% which was much higher than the 2.7% detected by ARMS-PCR with statistically different.
It showed that NGS could direct the drug use for the resistant patients. However, some rare loci could be detected by NGS but the importance and directed meaning are still unknown and the number of rare mutations is rare too. Further research on new biomarkers and technique is still needed for early diagnosis, directing drug use and assessing the therapy prognosis.
通过比较扩增受阻突变系统 PCR（ARMS-PCR）和下一代测序（NGS）在非小细胞肺癌（NSCLC）中靶向基因突变的检测率和类型，解释了非小细胞肺癌检测的特点和应用优势，为临床医生有效选择相应的检测方法提供了依据。
从 2016 年 1 月至 2020 年 10 月，选择重庆医科大学第一附属医院的肺癌靶向基因病例。选择了 4467 例病例，经病理活检诊断为 NSCLC。样本来源包括手术切除、支气管镜活检、转移活检、血液、痰液、胸腔积液细胞学。其中，3665 例采用 ARMS-PCR 技术检测，802 例采用 NGS 技术检测。比较了 ARMS-PCR 和 NGS 技术在不同 NSCLC 样本中 EGFR 基因突变（包括外显子 18、19、20、21 等）的检测率和类型。
ARMS-PCR 检测 EGFR 基因总突变率为 47.6%，NGS 检测为 42.4%，两种方法检测 EGFR 基因总突变率差异有统计学意义（P<0.001）。在不同外显子中，两种方法检测 EGFR 突变率不同。ARMS-PCR 检测外显子 19 的突变率明显高于 NGS 检测，而 ARMS-PCR 检测外显子 20 和 21 的突变率明显低于 NGS 检测。此外，NGS 检测的多重突变率为 16.3%，明显高于 ARMS-PCR 的 2.7%，差异有统计学意义。
NGS 可以为耐药患者提供药物指导。然而，NGS 可以检测到一些罕见的突变，但这些突变的重要性和指导意义尚不清楚，而且罕见突变的数量也很少。需要进一步研究新的生物标志物和技术，以实现早期诊断、药物指导和评估治疗预后。
