微滴数字PCR与ARMS法检测非小细胞肺癌EGFR和EML4-ALK基因的结果比较  

Comparison of EGFR and EML4-ALK genes detected by microdrop digital PCR and ARMS in non-small-cell lung cancer

作　　者：李秀忠[1] 郭雅琪 董辉 董洁 赵倩颖 赵志军 LI Xiuzhong;GUO Yaqi;DONG Hui;DONG Jie;ZHAO Qianying;ZHAO Zhijun(Department of Respiratory and Critical Disease Medicine,General Hospital of Ningxia Medical University,Yinchuan 750004,China;Medical Experiment Center,General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院呼吸与危重症医学科,宁夏银川750004 [2]宁夏医科大学总医院医学实验中心,宁夏银川750004

出　　处：《宁夏医学杂志》2020年第5期402-405,共4页Ningxia Medical Journal

基　　金：宁夏科技支撑计划资助项目(2015-26);宁夏医科大学科学研究基金资助项目(XM2018090)。

摘　　要：目的探讨微滴数字PCR技术在非小细胞肺癌(NSCLC)表皮生长因子受体(EGFR)基因与棘皮动物微管相关类蛋白4/间变性淋巴瘤激酶(EML4-ALK)融合基因联合检测中的应用价值。方法应用突变扩增阻滞系统(ARMS)法和微滴数字PCR(ddPCR)法同时检测73例NSCLC患者石蜡组织样本中EGFR基因和EML4-ALK融合基因的表达情况,并分析这两种检测方法的一致性。结果在73例NSCLC标本中,ARMS法检测EGFR基因的总突变率为43.84%(32/73),ddPCR法的总突变率为49.32%(36/73),两种检测方法具有高度一致性(K值=0.890,P<0.05)。在73例NSCLC标本中,ARMS法和ddPCR法检测EML4-ALK融合基因的总突变率均为5.48%(4/73),两种检测方法结果完全一致。有6例标本两种检测方法的结果不相符,其中2例标本ARMS法检测为阴性,ddPCR法检测为Exon 18-G719X位点阳性;2例标本ARMS法检测为阴性,ddPCR法检测为Exon 20-T790M位点阳性;1例标本ARMS法检测为Exon 19-del阳性,ddPCR法检测为19-del+T790M双重突变阳性;1例标本ARMS法检测为Exon 21-L858R阳性,ddPCR法检测为L858R+T790M双重突变阳性。EML4-ALK融合基因在两种检测方法中突变率均为5.48%(4/73)。两种方法均未检出EGFR和EML4-ALK基因的共存突变。结论 ddPCR技术在EGFR和EML4-ALK基因的联合检测方面具有良好的临床应用价值。Objective To explore the application value of microdrop digital PCR in the combined detection of EGFR gene and EML4-ALK fusion gene in non-small cell lung cancer.Methods The expression of EGFR gene and ALK fusion gene in paraffin tissue samples of 73 NSCLC patients were detected simultaneously by ARMS method and microdrop digital PCR method,and the consistency of the two detection methods were analyzed.Results In 73 NSCLC samples,the total mutation rate of EGFR detected by ARMS method was 43.84%(32/73),and the total mutation rate of ddPCR method was 49.32%(36/73).The two detection methods were highly consistent(κ=0.890,P<0.05);In 73 NSCLC specimen,the total mutation rate of EML4-ALK fusion gene detected by ARMS and ddPCR was 5.48%(4/73),and the results of the two detection methods were completely consistent;In 6 cases,the results of the two detection methods were inconsistent,among which the ARMS method was negative in two cases and the ddPCR method was positive at Exon 18-G719X.The ARMS test of the two samples was negative,and the ddPCR test was positive at the Exon 20-T790M site.One specimen was tested positive for Exon 19-del by ARMS method,and positive for double-mutation of 19-del+T790M by ddPCR method.One specimen was positive for Exon 21-L858R detected by ARMS method,and positive for L858R+T790M detected by ddPCR.The results of EML4-ALK fusion gene in the two detection methods were completely consistent,with a mutation rate of 5.48%(4/73).Coexisting mutations of EGFR and EML4-ALK gene were not detected in either method.Conclusion The ddPCR has a good clinical application value in the combined detection of EGFR and EML4-ALK gene.

关 键 词：EGFR基因 EML4-ALK融合基因 非小细胞肺癌 突变扩增阻滞系统 

分 类 号：R734.4[医药卫生—肿瘤]