High Resolution Melting Analysis Of KRAS And EGFR Mutation In Colorectal Cancer

Objective:To study the feasibility of high resolution melting (HRM) of KRAS and EGFR gene mutation in colorectal cancer. The effective and feasibility method to detect the polymorphism of KRAS and EGFR would be beneficial to prevent and diagnose the colorectal cancer, and would be hopeful to invent the new treatments.Methods:In this study, the70patients with clinically diagnosed colorectal cancer were selected from Xiangya hospital of Central South University and Provincial Tumor Hospital from September2012to December2012. Genomic DNA was extracted and purified from blood samples which from the patients donated. Designed the primers of PCR and amplified the KRAS exon2, KRAS exon3, EGFR exon19and EGFR exon21respectively. The genotype of KRAS and EGFR were detected using HRM, and meanwhile checked the result of HRM using sequencing technology.Result:There are18cases harboured KRAS exon2mutations by HRM, the types of mutations are G12D(6), G12C(9), G12V(3), but there are only17case harboured mutations were detected by sequencing. Using the HRM and the sequencing found that nobody was confirmed the presence of KRAS exon3and EGFR exon21, but there are3samples were positive for EGFR exon19mutations.Conclusion:The technology of high resolution melting could get the same results as sequencing. HRM is a sensitive and precise in-tube methodology to detect the mutations in clinical samples, so it is well worth applying in clinical study and result in substantial time and cost savings and may find a new method to cure the patient.