高分辨率熔解曲线分析法检测非小细胞肺癌KRAS和BRAF基因突变

Detection of KRAS and BRAF mutations in non-small cell lung cancer by high resolution melting analysis

目的探讨高分辨率熔解曲线分析(HRM)法检测非小细胞肺癌(NSCLC)中KRAS和BRAF基因突变用于临床检测的可行性。方法用HRM法检测64例NSCLC患者KRAS基因第2外显子和BRAF基因第15外显子的突变情况,用直接测序法对结果进行验证。结果 HRM法检测结果表明有9例NSCLC患者发生KRAS基因突变(14.06%)、4例发生BRAF基因突变(6.25%),直接测序法证实两法的结果完全一致;共检测出4种KRAS基因突变类型,G12C(GGT>TGT)的突变率最高(44.4%),BRAF基因突变型均为V600E。结论用HRM法检测临床样本KRAS和BRAF基因突变,具有操作简便、结果准确、成本低的优点,适用于临床检测。

Objective To evaluate the feasibility of detecting KRAS and BRAF mutations in the patients with non-small cell lung cancer （NSCLC） by high resolution melting curve analysis （HRM）. Methods The mutations of KRAS exon 2 and BRAF exon 15 in 64 pa- tients with NSCLC were detected by HRM analysis and direct sequencing. Results KRAS exon 2 mutations were detectable in 9 pa- tients （ 14.1% ）, and BRAF exon 15 mutations were detectable in 4 patients（6.25% ） by both the two methods. KRAS mutations were comprised of 4 different mutant alleles. G12C showed the highest mutation rate （44.4%）. The mutations type of BRAF in exon 15 were all V600E. The concordant rate between the two methods was 100%. Conclusion Compared with direct sequencing, HRM a- nalysis for detecting KRAS and BRAF should be an accurate, convenient, efficient and economical assay which is suitable for clinical application.