Comparison of High-Resolution Melting Analysis, Sanger Sequencing and ARMS for KRAS Mutation Detection in Metastatic Colorectal Cancer

Background: 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. Methods: 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. Results: 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. Conclusions: 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.