Sanger测序法和突变扩增阻滞系统法检测非小细胞肺癌EGFR基因19、21号外显子突变的临床价值比较

Detection of EGFR gene 19 and 21 exons mutations in non-small cell lung cancer by Sanger sequencing versus amplification refractory mutation system assay

目的比较Sanger测序法和突变扩增阻滞系统(ARMS)法检测非小细胞肺癌(NSCLC)EGFR基因19、21号外显子突变的临床价值。方法收集经病理组织学确诊的NSCLC患者肺部原发或转移癌标本102例,其中石蜡包埋组织75例,病理活检标本27例;采用Sanger测序法和ARMS法检测上述标本EGFR基因19、21号外显子突变情况,分析其与患者临床病理学特征的关系。结果 ARMS法突变检出率为48.0%(49/102),高于Sanger测序法的32.3%(31/96),两者比较差异有统计学意义(P<0.05)。病理活检组织ARMS法突变检出率为57.1%(12/21),明显高于Sanger测序法的23.8%(5/21),差异有统计学意义(P<0.05);石蜡包埋组织ARMS法和Sanger测序法突变检出率分别为49.3%(37/75)和34.7%(26/75),两者比较差异无统计学意义(P>0.05)。女性患者EGFR基因突变检出率68.0%高于男性患者的28.8%,未吸烟患者突变检出率65.5%高于吸烟患者的27.7%,腺癌患者突变检出率62.3%高于鳞癌患者的26.8%,差异均有统计学意义(均P<0.05);但EGFR基因突变检出率和有无淋巴结转移及年龄比较差异均无统计学意义(均P>0.05)。结论 EGFR基因19、21号外显子突变好发于女性、未吸烟患者和腺癌患者,Sanger测序法对大组织样本及未知突变检测更有优势,ARMS法对病理活检、微小样本及要求灵敏度高的样本检测更为适合,结合2种方法检测结果更为全面可靠。

Objective To compare Sanger sequencing and amplification refractory mutation system(ARMS)assay indetection of EGFR gene19and21exons mutations in non-small cell lung cancer(NSCLC).Methods One hundred and twoNSCLC specimens,including75paraffin tissue samples and27biopsy specimens were collected.Mutations of EGFR gene19and21exons were detected by Sanger sequencing and ARMS assay.The association of gene mutations with clinicopathologicalfeatures of NSCLC was analyzed.Results The mutation rates of ARMS assay and Sanger sequencing were48.0%(49/102)and32.3%(31/96),respectively(P<0.05).In biopsy tissue samples,the mutation rate detected by ARMS(57.1%,12/21)wassignificantly higher than that by Sanger sequencing(23.8%,5/21,P<0.05),however,there was no significant difference inmutation rate of paraffin embedded specimens between two methods(49.3%,37/75vs34.7%,26/75,P>0.05).According toARMS results,the EGFR mutation rate was higer in female patients(68.0%)than in male patients(28.8%),in non-smoking patients(65.5%)higher than that in smoking patients(27.7%),in adenocarcinomas(62.3%)higher than that in squamous cell carcinomas(26.8%)(all P<0.05).The EGFR mutation rate was not correlated with the age and lymphatic metastasis of patients(all P>0.05).Conclusion The mutation rates of EGFR19and21exons were higher in NSCLC patients,especially in non-smoking,female andadenocarinoma patients.Sanger sequencing method has more advantages for large tissue samples,and the ARMS method ismore suitable for biopsy and small samples.Therefore,the combination of two methods may provide more reliable andcomprehensive test results.