Zhang Haiyan, Zhang Xiaoming, Wang Jie, Xian Jiang, Chen Xiaodong, Zhang Wei
Clin Lab. 2015;61(3-4):435-9. doi: 10.7754/clin.lab.2014.140923.
Treatment of metastatic colon carcinoma with the anti-epidermal growth factor receptor antibody cetuximab/panitumumab is reported to be ineffective in KRAS-mutant tumors; therefore, it is necessary to perform KRAS mutation analysis before cetuximab or panitumumab treatment is initiated.
This study was designed to compare and evaluate the efficacy of three different methodologies--high resolution melting (HRM), Sanger sequencing, and Amplification Refractory Mutation System (ARMS)--for KRAS mutation detection in a clinical setting.
In total, 55 samples from patients with metastatic colorectal cancer were analyzed. Compared to Sanger sequencing, good consistency was found between the results of the ARMS (Kappa = 0.839) and HRM (Kappa = 0.839). The sensitivities of the methods were compared after a consensus was reached: if two of the three methodologies showed a similar result, it was considered as the consensus result. The frequency of KRAS mutations in our population was 34.5%, and discordant findings were observed in five samples. No significant difference in sensitivity was found among the three methodologies.
From the results, we can conclude that after careful in-laboratory validation, HRM is a good alternative to the ARMS and Sanger sequencing for KRAS mutation testing.
据报道，用抗表皮生长因子受体抗体西妥昔单抗/帕尼单抗治疗转移性结肠癌对KRAS突变型肿瘤无效；因此，在开始西妥昔单抗或帕尼单抗治疗前进行KRAS突变分析很有必要。
本研究旨在比较和评估三种不同方法——高分辨率熔解曲线分析（HRM）、桑格测序法和扩增阻滞突变系统（ARMS）——在临床环境中检测KRAS突变的疗效。
总共分析了55例转移性结直肠癌患者的样本。与桑格测序法相比，ARMS（Kappa = 0.839）和HRM（Kappa = 0.839）的结果之间具有良好的一致性。在达成共识后比较了这些方法的敏感性：如果三种方法中的两种显示出相似的结果，则将其视为共识结果。我们研究人群中KRAS突变的频率为34.5%，在五个样本中观察到不一致的结果。三种方法在敏感性上没有发现显著差异。
从结果来看，我们可以得出结论，经过仔细的实验室验证后，HRM是用于KRAS突变检测的ARMS和桑格测序法的良好替代方法。
