高分辨率熔解曲线分析法检测结直肠癌KRAS基因突变

Detection of KRAS Mutations in Colorectal Cancer by High Resolu tion Melting Analysis

目的:采用高分辨率熔解曲线分析法检测结直肠癌中KRAS基因突变,探讨其用于临床检测的可行性。方法:首先用高分辨率熔解曲线分析法检测64例结直肠癌患者KRAS基因第2外显子的突变情况,再用直接测序法对结果进行验证。结果:通过高分辨率熔解曲线分析法检测,发现有23例KRAS基因突变(35.9%),经直接测序法验证,证实所有患者的突变情况与高分辨率熔解曲线法的结果完全一致;共检测出6种KRAS突变类型,G12D(GGT>GAT)的突变率最高(47.8%),G12D、G12V和G13D等3种常见突变型占总突变数的78.3%。结论:与直接测序法相比,应用高分辨率熔解曲线分析法检测KRAS基因突变具有操作简单快捷、结果准确、成本低的优点,适合应用于临床检测。

Objective： To evaluate the feasibility of detecting the KRAS mutations in colorectal cancer patients by high resolution melting curve analysis.Methods： KRAS exon 2 mutations were performed on 64 patients with colorectal cancer by high resolution melting analysis and direct sequencing.Results： KRAS exon 2 mutations were detected in a total of 23 patients（35.9%） with the two methods combined,comprising 6 different mutant alleles.Three common mutations G12D,G12V and G13D accounted for 78.3% of all cases,mutations of G12D was the highest（47.8%）.The concordant rate between the two methods was 100%.Conclusion： Compared with direct se quencing,high resolution melting curve analysis is accurate,convenient,efficient and economical,which is suitable for clinic test.