环介导等温扩增法检测转基因玉米MON88017品系  

A Specific Loop-Mediated Isothermal Amplification Method for the Detection of Transgenic Maize Event MON88017

作　　者：马丹[1] 张沫琦 魏海燕[1] 魏咏新[1] 徐蕾蕊[1] 曾静[1] 

机构地区：[1]北京出入境检验检疫局,北京100026

出　　处：《检验检疫学刊》2017年第2期1-5,共5页Journal of Inspection and Quarantine

基　　金：国家质检总局科技计划项目(2015IK341)

摘　　要：建立了转基因玉米MON88017品系环介导等温扩增法检测方法。针对MON88017玉米基因组与外源基因结合处序列,设计5条特异引物,并对反应条件、特异性、灵敏度、稳定性、结果判断方法等参数进行研究,同时与实时荧光PCR检测方法进行比较,最终进行了5家同类型实验室的协同验证。结果表明本研究所建立的MON88017环介导等温扩增检测方法具有良好的特异性,其最低检出限为0.5%,在特异性、灵敏度和检测范围等指标上均优于传统的PCR技术,并且不依赖昂贵的大型设备,可以实现现场快速检测,检测成本与检测所需时间远低于实时荧光PCR。该方法具有快速、简便的优点,能满足实际工作要求。In this study,the LAMP detection method for transgenic maize MON88017 was established. Five pairs of specific primers cover the MON88017 maize genome junction with exogenous sequences region were designed for LAMP reaction. The specificity,sensitivity and stability of the LAMP method were analyzed. The reaction conditions were optimized. And also the specificity,sensitivity and stability of the LAMP method were compared with real-time flurosecence PCR method （RT-PCR）. An inter-laboratory co-verification study was conducted. The results show that the establishment of MON88017 loop-mediated isothermal amplification detection method has good specificity,the minimum detection limit of 0.5%. The technology in the specificity,sensitivity and detection range of other indicators better than traditional PCR techniques,does not rely on expensive equipment,you can achieve on-site rapid testing,test time and inspection costs far less than the required for real-time fluorescence PCR.LAMP detection method of genetically modified maize line MON88017 is simple,raoid and soecific,to meet the actual testing reauirernents.

关 键 词：转基因玉米 环介导等温扩增法 MON88017 

分 类 号：TS207[轻工技术与工程—食品科学]