胶质瘤表皮生长因子受体基因突变检测方法的对比研究

Comparative study of mutation detection method in glioma epidermal growth factor receptor gene

目的分析普通聚合酶链式反应(PCR)联合直接测序法与实时聚合酶链式反应-高分辨率融解(qPCR-HRM)曲线分析技术对胶质瘤表皮生长因子受体(EGFR)基因突变的检测效果。方法用普通PCR联合直接测序法与qPCR-HRM曲线分析技术检测胶质瘤患者EGFR基因突变类型,检测结果比较采用χ2检验。结果普通PCR联合直接测序法与qPCR-HRM曲线分析技术对胶质瘤患者肿瘤组织EGFR突变的检测结果一致性较高,达90.91%,两者间差异无统计学意义(P>0.05)。结论 qPCR-HRM曲线分析技术操作较普通PCR联合直接测序法简便,实验时间较短,降低了交叉污染的风险,通过HRM曲线分析检测结果可以实现对样品基因型的判定。

Objective To analyse mutation detection effect of the ordinary polymerase chain reaction（PCR） method with direct sequencing method and real-time polymerase chain reaction-high resolution melting curve analysis techniques（qPCR-HRM） curve analysis technology on the detection of glioma epidermal growth factor receptor（EGFR） gene.Methods Ordinary PCR with sequencing method and qPCR-HRM curve analysis technical were used to detect EGFR gene mutation type in patients with glioma,the chi test was used for results comparison.Results The consistency of detection results was up to 90.91% between ordinary PCR with direct sequencing method and qPCR-HRM curve analysis technology on tumor tissue in patients with glioma EGFR mutation,the difference was statistically significant（P0.05）.Conclusion Compare to ordinary PCR with direct sequencing method,the operation of qPCR-HRM curve analysis technical might more simple,the experimental time might shorter,the risk of cross contamination might less,and its test results could identify the samples of genotype.