高分辨率熔解曲线分析技术在检测非小细胞肺癌组织EGFR基因突变中的应用

Detecting EGFR mutations in non-small cell lung carcinoma patients using high resolution melting curve analysis

目的:探索高分辨率熔解曲线(high resolution melting,HRM)分析技术在检测肺癌组织表皮生长因子受体(epidermal grouthfactor receptor,EGFR)基因突变中的应用。方法:利用HRM技术,进行人肺癌组织EGFR基因外显子19和21突变筛查,并做测序验证。结果:外显子19的HRM分型结果显示,52例样品中有3种类型熔解曲线,45例样品为A型,5例为B型,2例为C型。外显子21的分析结果显示有2种熔解曲线,49例样品为N型,3例样品为M型。测序结果显示,外显子19 A型熔解曲线代表外显子19的野生型,B型熔解曲线代表杂合性缺失delE746-A750(c.2235-2249del),C型熔解曲线代表杂合性缺失delL747-S752(c.2240-2257del)。外显子21 N型熔解曲线代表野生型,M型熔解曲线代表杂合性L858R(c.2573T-G),HRM分析结果与测序结果完全一致。结论:本研究表明HRM分析技术能准确检测EGFR基因突变,是一种适合肺癌组织中EGFR基因突变筛查的方法。

OBJECTIVE： To evaluate the feasibility and sensitivity of detecting the epidermal growth factor receptor（EGFR） mutation in non-small cell lung carcinoma（NSCLC） patients using high resolution melting（HRM） curve analysis.METHODS： HRM was applied to detect the mutations of EGFR exon 19 and 21 in NSCLC patients,the results were then confirmed by direct sequencing.RESULTS： Three melting curves were obtained by HRM anlysis for EGFR exon 19 and two melting curves for exon 21.Type A（45/52）,B（5/52）,C（2/52） melting curves represent wild type,heterozygous delE746-A750 and delL747-S752 of exon 19,respectively.Type N（49/52） and M（3/52） melting curves represent wild type and heterozygous L858R of exon 21.Results of HRM were consistent with those of direct sequenc.CONCLUSION： HRM is an accurate technique to detect EGFR mutations.It is also a low-cost test which is suitable for screening.