Development of a Rapid Event-Specific Loop-Mediated Isothermal Amplification Detection Method for Genetically Modified Maize NK603

摘要

In the present study, a loop-mediated isothermal amplification (LAMP) system was developed for detecting NK603 maize, and flanking sequence in NK603 maize was used as the target template to design event specific LAMP primers. Different kinds of genetically modified (GM) or non GM crops were selected to ensure the specificity of detection and the results of the non-target samples were negative, indicating that the primer set for NK603 maize had good levels of specificity. The limit of detection (LOD) of the LAMP system was about 50 copies of the maize haploid genome. Moreover, the amplified reaction could be accomplished in about 1 h, and the LAMP amplification products tend to be far longer than conventional PCR products and thus easier to be visible to naked eyes via end point analysis. Therefore, the LAMP method developed by this study is visual, faster, and more sensitive, and does not need special equipment compared to traditional PCR technique, which is very useful for detecting transgenic maize aimed at screening the growing transgenic crops in the fields and detecting GM ingredients in imported and domestic foods.