AMSS-PCR在肺癌突变基因检测中的价值研究  

Applicable Value of AMSS-PCR in Lung Cancer Gene Mutation Detection

作　　者：金柯[1] 谢绚[1] 潘越江[1] 王科喜 陈柏深[1] 吴多光[1] 沈卓坚[1] 王铭辉[1] 张惠忠[1] Ke JIN;Xuan XIE;Yuejiang PAN;Kexi WANG;Baishen CHEN;Duoguang WU;Zhuojian SHEN;Minghui WANG;Huizhong ZHANG(Department of Thoracic Surgery,Sun Yat-Sen Memorial Hospital of Sun Yat-Sen University,Guangzhou 510120,China)

机构地区：[1]中山大学孙逸仙纪念医院胸外科,广州510120

出　　处：《中国肺癌杂志》2018年第11期815-820,共6页Chinese Journal of Lung Cancer

摘　　要：背景与目的肺癌驱动基因检测具有重要意义,目前检测方法多样,临床适用性有差异。本研究旨在比较基于扩增阻碍突变系统-聚合酶链反应(Amplification Refractory Mutation System-polymerase chain reaction,ARMS-PCR)技术的试剂盒与一代测序及ARMS-qPCR检测肺癌突变基因的敏感性和特异性,探究突变位点特异扩增法(Amplification Mutation Specific System, AMSS)-PCR技术在肺癌突变基因检测中的应用价值。方法对前期已行ARMS-PCR检测的肿瘤标本进行一代测序及试剂盒检测,比较各种方法的检测结果,并对检测结果进行统计学分析。结果本研究共收集了309例肺癌标本。试剂盒与一代测序符合率97.41%,ARMS-PCR的符合率97.73%。试剂盒与一代测序、试剂盒与实时定量聚合酶链反应(quantitative real-time polymerase chain reaction, qPCR)、qPCR与一代测序一致性检验的Kappa值分别为0.946、0.953、0.913。试剂盒以一代测序为参照的受试者工作特征曲线(receiver operatingcharacteristiccurve,ROC)曲线下面积为0.976,以qPCR为参照的ROC曲线下面积为0.975。结论 AMSSqPCR技术能够有效检测肺癌突变基因,具有较好的临床应用价值。Background and objective The detection of driver oncogenes of lung cancer is of great importance. There are various gene detection techniques nowadays which are different from each other.We carried out this study to inves- tigate the specificity and sensitivity of assay panels based on an Amplification Refractory Mutation System-polymerase chain reaction (ARMS-PCR)technique of Amplification Mutation Specific System (AMSS)in detection of lung cancer gene muta- tion.To estimate the applicable value ofassaypanels in cliuical settings.Methods We collected cancer tissue specimens or fluid specimens from 309patients.Mutation results were presented for those samples previously detected by ARMS-PCR.In com- parison,we carried out AMSS-P CR using (epidermal growth factor receptor,EGFR)assay panel and Six-AlLiance assay panel as well as Sanger sequencing.Software SPSS 22.0(SPSS IBM)was used for statistical analysis.Results The rates of consistency between the results by assay pands and Sanger sequencing or ARMS-PCK were 97.41%and 97.73%,respectively.Besides, EGFR assay panel had higher consistency rates with other detection methods than Six-AUiance assay panel.As for consistency test,the Kappa values of assay panels with Sanger sequencing,assay panels with ARMS-PCR,and ARMS-PCK with Sanger se- quencing were 0.946,0.953,and 0.913,respectively.The receiver operating characteristic curve (ROC)area under curve (AUC) of assay panels was 0.976referring to Sanger sequencing,and 0.975as ARMS-PCKwas referred to.Conclusion AMsS-PCR can make an optimal cancer gene mutation detection method for cliuicalsettings.

关 键 词：一代测序 突变位点特异扩增法 扩增阻碍突变系统 实时定量聚合酶链式反应 肺癌突变基因检测 

分 类 号：R734.2[医药卫生—肿瘤] R730.43[医药卫生—临床医学]