变性高效液相色谱法检测非小细胞肺癌患者外周血及肿瘤组织表皮生长因子受体突变

目的 建立变性高效液相色谱(DHPLC)法检测晚期非小细胞肺癌(NSCLC)患者外周血与肿瘤组织中表皮生长因子受体(EGFR)基因外显子19和21突变的技术平台.方法 同时收集2004年4月至2007年1月在北京肿瘤医院接受治疗的230例晚期NSCLC患者的血浆和肿瘤组织标本,采用DHPLC法检测EGFR外显子19和21突变并用测序法验证,应用x2检验分析EGFR突变与患者临床病理特征的关系,以调整比值比(OR)及95%可信区间(CJ)表示相对危险度,所有统计检验均为舣侧概率检验.分析在血浆和肿瘤组织标本中基因突变的一致性及DHPLC法与测序法的一致性.结果 230例血浆标本中EGFR突变率为34.3%(79/230),肿瘤组织中为33.5%(77/230).血浆和肿瘤组织突变阳性检出一致性为79.7%(63/79,k值为0.74).与测序法相比,DHPLC法的灵敏度和特异度分别为96.9%和91.9%(k值为0.88).肿瘤组织及外周血中EGFR突变与病理类型(OR=3.38,95%CI 1.81～6.36,P＜0.05)、吸烟史(OR=1.61,95% CI 1.13～2.28,P＜0.05)相关,而与年龄、性别、病理分期无明显相关性.结论 晚期NSCLC患者血浆EGFR突变率与肿瘤组织的突变率具有高度一致性,DHPLC法可作为EGFR突变检测的初筛方法.

The detection by denaturing high performance liquid chromatography of epidermal growth factor receptor mutation in tissue and peripheral blood from patients with advanced non-small cell lung cancer

Objective To study the application of denaturing high performance liquid chromatography (DHPLC) as a screening tool in detecting plasma and matched tissue epidermal growth factor receptor (EGFR) mutations for advanced non-small-cell lung cancer ( NSCLC).MethodsPlasma DNA samples and matched tumors from 230 cases of NSCLC were analyzed for EGFR mutations in exons 19 and 21 using DHPLC.The mutations in the pasma samples and the matched tumors were compared,and the association between EGFR mutations and the clinicopathological features were evaluated.Results Mutation of EGFR was found by DHPLC to be 33.5% (77/230) in tissues and 34.3% (79/230) in matched peripheral blood samples.Consistency of EGFR mutation status between tissues and matched plasma DNA was confirmed (k is 0.74,P ＜ 0.01 ).The sensitivity and specificity of DHPLC for detecting EGFR mutation were 96.9% and 91.9%,respectively(K is 0.88).EGFR mutations in both tissue and blood was correlated with histology type(OR = 3.38,95% CI 1.81-6.36,P ＜0.05 ) and smoking status( OR = 1.61,95%CI 1.13-2.28,P ＜0.05),but no association with age,sex and stage was found(P ＞0.05).Conclusion The detection of EGFR mutation is highly consistent in tissues and in plasma DNA samples.DHPLC may serve as a preliminary screening tool for detecting EGFR mutations.