直接测序法和qPCR-HRM法检测NSCLC组织EGFR突变的比较研究

A comparative study on detecting EGFR mutation in NSCLC tissue by qPCR-HRM and DNA sequencing

目的比较直接测序法和实时聚合酶链反应-高分辨率融解曲线技术(q PCR-HRM)检测非小细胞肺癌(NSCLC)表皮生长因子受体(EGFR)基因突变的差异,探讨适用于临床检测EGFR基因突变的方法,为NSCLC患者的靶向治疗提供理论依据。方法共收集NSCLC患者组织标本52例,分别采用q PCR-HRM法和直接测序法进行EGFR基因突变检测,采用χ2检验对检测结果进行统计学比较。结果 52例NSCLC患者组织标本中,直接测序法检测的EGFR基因突变率为30.8%(16/52),q PCR-HRM法检测的EGFR基因突变率为32.7%(17/52),两种方法检测结果相比较,差异无统计学意义(χ2=0.04,P>0.05)。结论与直接测序法相比较,q PCR-HRM法是一种操作简便、费用低、速度快、灵敏度高的检测方法,对于临床筛选适合EGFR-TKI治疗的NSCLC患者亚群,预测EGFR-TKI疗效都具有重要意义,值得临床进一步推广应用。

Objective To compare the utility of Quantitative PCR high-resolution melting（ qPCR-HRM） curve analysis and DNA sequencing in the detection of EGFR mutations in NSCLC tissues,investigate the detection methods of EGFR gene mutation which is more suitable for clinical diagnosis,and provide evident for the targeted therapy of patients with NSCLC. Methods EGFR mutation of tumor tissue was detected both by qPCR-HRM method and DNA sequencing in 52 patients with NSCLC. Chi-square test was used to analyze the consistency. Results EGFR mutation rate was 30. 8%（ 16 /52） by qPCR-HRM method and 32. 7%（ 17 /52） by DNA sequence method,respectively; there was no significant difference between the results of the two methods.（ χ^2= 0. 04,P〉0. 05）. Conclusion As compared with DNA sequencing,qPCR-HRM method was simple,cheap,rapid and sensitive,which has the vital significance for screened EGFR-TKI treatment subgroups of patients with NSCLC and predicted the curative effect of EGFR-TKI,and is applicable in the clinical setting.