高分辨率熔解曲线分析技术检测EGFR基因突变方法的建立及其初步临床应用

目的建立高分辨率熔解曲线(high resolution melting,HRM)分析技术检测表皮生长因子受体(epidermal growth factor receptor,EGFR)基因突变的方法,并探讨其的临床应用价值。方法建立HRM技术检测EGFR基因突变方法,并用其检测200例非小细胞肺癌患者肿瘤石蜡包埋标本,并与测序法的结果进行比较。结果所建HRM检测方法 Tm与ct值的CV值均较小,重复性好。HRM法检测标本EGFR基因突变的结果与测序法相比,突变总检出率分别为40.0%和37.0%,敏感性为100%,特异性>95%。结论 HRM法检测EGFR基因突变,敏感性高,特异性强,重复性好,操作简便,节约时间,成本低,适用于临床。

Establishment and primary clinical application of detecting EGFR mutations by high resolution melting analysis

Objective To establish a method for detection of EGFR mutations by high resolution melting （HRM） analysis and to evaluate its clinical application value. Methods 200 formalin-fixed paraffin-embedded tissues of NSCLC patients were detected by HRM which have been established. The results of HRM analysis were confirmed by direct DNA sequencing. Results According to the repeatability analysis, it was found that the value of Coefficient of Variance （CV） of Tm and ct was very small. The total EGFR mutation rates were 37.0% detected by direct sequencing and 40.0% detected by HRM. The sensitivity of HRM assay was 100% , the specificity of HRM assay was above 95%. Conclusions The method for detection of EGFR mutations by HRM analysis is appropriate for detecting EGFR mutations by HRM analysis. It possesses high specificity, good sensitivity, strong reproducibility, simple operation, saving-time and low cost qualities.