转Bt基因作物恒温扩增快速检测技术

Rapid Detection of Transgenic Bt Crops by Isothermal Nucleic Amplification

[目的]建立一种快速检测转Bt基因作物的方法。[方法]利用Bt的Cry1ab/ac基因序列设计特异性引物,建立LAMP检测方法以及优化反应体系,并进行LAMP的特异性和灵敏度试验。[结果]成功建立转Bt基因作物的LAMP检测方法,得到最优的反应体系。在特异性试验中,转Bt基因作物基因组DNA均呈阳性,而非转基因作物、转Bar基因作物、转CPTI基因作物、转EPSPS基因作物等均为阴性;在灵敏度试验中,转Bt基因作物的最低检测限为10个拷贝。[结论]LAMP方法具有很高的特异性和灵敏度,可用于转Bt基因作物的现场快速检测。

[Objectives] To establish a specific,sensitive and rapid method for detecting transgenic Bt crops.[Methods] Cry1ab/ac gene was used for the design of primers,the reaction conditions were optimized and then LAMP method was established.Specificity and sensitivity evaluations also included in the experiment.[Results] LAMP method was established successfully,it was proved to be a high specific method which could detect all transgenic Bt crops,while results of detection of non-transgenic crops,transgenic Bar crops,transgenic CPTI crops,transgenic EPSPS crops,etc.were all negative.The detection limit of LAMP was 10copies/tube for transgenic Bt crops.[Conclusions] The established LAMP was a specific and sensitive method for detecting transgenic Bt crops and it can be used for Rapid detection of transgenic Bt crops.