LAMP实时浊度法快速检测转基因玉米MON810  

A Real-time Turbidimeter-based Loopmediated Isothermal Amplification Assay for Rapid Detection of Genetically Modified Maize MON810

作　　者：王小玉[1,2] 邝筱珊[2] 胡松楠[2] 余以刚[1] 成晓维[2] 冯家望[2] 张璜 肖性龙[1] 

机构地区：[1]华南理工大学轻工与食品学院,广东广州510640 [2]珠海出入境检验检疫局技术中心,广东珠海519015 [3]广州迪澳生物科技有限公司,广东广州510663

出　　处：《现代食品科技》2013年第12期3002-3005,3069,共5页Modern Food Science and Technology

基　　金：国家质检总局科研基金项目(2011IK255);国家自然科学基金项目(31101279);教育部高等学校博士学科点专项科研基金项目(20110172120034)

摘　　要：LAMP实时浊度法是采用环介导等温扩增(Loop Mediated Isothermal Amplification,LAMP)技术通过实时浊度仪实时检测反应过程中所产生的白色沉淀,从而实现对扩增全过程的监控,弥补了显色法只能观看反应终点的缺陷,使引物筛选和反应体系的优化有数据可依。本研究以转基因玉米MON810为研究对象,针对外源基因苏云金芽孢杆菌杀虫毒蛋白CryIA(b)与内源基因边界序列设计6条特异性引物,通过实时浊度法在63℃的恒温条件下完成检测,对检测的灵敏度、特异性、稳定性进行了评价。建立了转基因玉米MON810的LAMP实时浊度检测方法,该方法最低检出限为0.5%,与LAMP显色法和实时荧光PCR法进行结果比对,符合率为100%,经评价具有特异性高、稳定性强、准确简便等优点,非常适合转基因玉米MON810的快速检测,有较好的应用价值。LAMP real-time turbidity method is based on the loop-mediated isothermal amplification, detecting a white precipitate generated during the reaction by a real-time turbidimeter, in order to monitor the whole process of amplification. This technique overcomes the defects of LAMP chromomeric method which could only monitor the end of reaction, therefore becomes more scientific for primers screening and the reaction optimization. Six specific primers were designed for the junction sequences of exogenous gene CryIA（b） and endogenous gene of genetically modified maize MON810. Detection conducted by real-time turbidity method at 63 ℃, the detection sensitivity, specificity and stability were verified. The results indicated that the lowest detection limit of this method was 0.5%, and the coincidence rate compared with the LAMP chromomeric method and the real-time PCR method was 100%, The method had high specificity, stability, accuracy and convenience which was suitable for the rapid detection of genetically modified maize MON810 and has good application value.

关 键 词：环介导等温扩增技术 实时浊度仪 转基因玉米 MON810 检测 

分 类 号：TS207.3[轻工技术与工程—食品科学]