核酸试纸条在检测转EPSPS基因作物中的应用

目的:建立快速、简便和特异的检测转EPSPS基因作物的方法。方法:针对转EPSPS基因作物外源基因cp4-EPSPS的8个区域,设计2对特异性引物和1对环引物,对反应体系中的MgSO4、内引物、环引物、甜菜碱、dNTP浓度和反应温度等条件分别进行优化,并用核酸试纸条对扩增产物进行检测,建立用于检测转EPSPS基因作物的环介导等温扩增方法(LAMP)。结果:用建立的LAMP方法检测转EPSPS基因作物时,在64℃恒温反应30 min,即可根据试纸条显色直接观察结果;该方法具有高度特异性,可检测到10个拷贝的EPSPS DNA。结论:LAMP方法可快速、灵敏、特异、经济地检测转EPSPS基因作物,在基层和实验室都具有良好的应用前景。

Application of Nucleic Acid Strips in the Detection of Transgenic EPSPS Crops

Objective：To develop a rapid,simple and sensitive loop-mediated isothermal amplification method for the detection of transgenic EPSPS crops.Methods：Two pairs of primers and a set of loop primer targeting the EPSPS gene were designed.The concentration of MgSO4,inner primer,loop primer,betaine,dNTP,and temperature were optimized.Using nucleic acid strips to detect the amplification products.Results：The amplification was carried out in a single tube at 64℃,and the amplification results were visualized according to the strips within 30 min.This test has a high specificity for transgenic EPSPS gene crops detection,and the minimum limit detected was 10 copies.Conclusion：The method described in this study was proved to be simple,sensitive,specific and rapid way for the detection of transgenic EPSPS crops.It has a potential application of both laboratorial and pen-side detection of transgenic EPSPS crops.