Cai Ling, Wang Weidong, Wang Fang, Zhang Rusi, Zhang Lanjun, Qi Huiwei, Gu Weiquan, Zhou Ningning
State Key Laboratory of Oncology in South China, Collaborative Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou 510060, China.
Department of Radiation Oncology, Sun Yat-sen University Cancer Center, Guangzhou 510060, China.
Transl Cancer Res. 2019 Nov;8(7):2564-2569. doi: 10.21037/tcr.2019.10.37.
Targeted therapy has been proven to be effective in lung cancer patients with specific driver gene mutations. At present, Sanger sequencing is still the gold standard in clinical practice to detect mutation, and amplification refractory mutation system PCR (ARMS-PCR) has become widely used due to its higher sensitivity and less limitation compared with Sanger sequencing. Mutation-selected amplification specific system PCR (MASS-PCR) is a novel gene detection technique with high specificity and sensitivity. This study aimed to compare the accuracy and sensitivity of ARMS-PCR and MASS-PCR and purposed to make an alternative choice in gene mutation detection in lung cancer.
A total of 293 formalin-fixed paraffin-embedded (FFPE) tissues were collected from 293 patients with lung cancer from 2017 to 2018. The sample mutation statuses were evaluated by ARMS-PCR and MASS-PCR. Sanger sequencing was also conducted to confirm the results further. The consistency of ARMS-PCR and MASS-PCR were analyzed, and receiver operating characteristic (ROC) curve was drawn to assess the sensitivity and specificity of MASS-PCR.
The consistency rate between the MASS-PCR and Sanger sequencing (kappa value =0.929) was higher than that between the MASS-PCR and ARMS-PCR (kappa value =0.821). There were 20 samples had inconsistent results among the three assays. For these samples, 11 positive samples were verified by the MASS-PCR and Sanger sequencing. Besides, 3 negative samples in Sanger sequencing were detected to be positive in MASS-PCR and ARMS-PCR. The ROC area under the curve (AUC) of assay panels was 0.930 referring to ARMS-PCR, and 0.967 as Sanger sequencing was referred to.
Our study demonstrated a higher accuracy and sensitivity of MASS-PCR than ARMS-PCR. Therefore, MASS-PCR could be used in clinical practice to detect gene mutations in lung cancer patients.
靶向治疗已被证明对具有特定驱动基因突变的肺癌患者有效。目前，桑格测序仍是临床实践中检测突变的金标准，而与桑格测序相比，扩增阻滞突变系统聚合酶链反应（ARMS-PCR）因其更高的灵敏度和更少的局限性而得到广泛应用。突变选择扩增特异性系统聚合酶链反应（MASS-PCR）是一种具有高特异性和灵敏度的新型基因检测技术。本研究旨在比较ARMS-PCR和MASS-PCR的准确性和灵敏度，并旨在为肺癌基因突变检测提供另一种选择。
收集2017年至2018年293例肺癌患者的293份福尔马林固定石蜡包埋（FFPE）组织。通过ARMS-PCR和MASS-PCR评估样本突变状态。还进行了桑格测序以进一步确认结果。分析ARMS-PCR和MASS-PCR的一致性，并绘制受试者工作特征（ROC）曲线以评估MASS-PCR的灵敏度和特异性。
MASS-PCR与桑格测序之间的一致性率（kappa值=0.929）高于MASS-PCR与ARMS-PCR之间的一致性率（kappa值=0.821）。三种检测方法中有20个样本结果不一致。对于这些样本，11个阳性样本经MASS-PCR和桑格测序验证。此外，桑格测序中的3个阴性样本在MASS-PCR和ARMS-PCR中检测为阳性。以ARMS-PCR为参照，检测组的曲线下面积（AUC）为0.930，以桑格测序为参照则为0.967。
我们的研究表明MASS-PCR比ARMS-PCR具有更高的准确性和灵敏度。因此，MASS-PCR可用于临床实践中检测肺癌患者的基因突变。
