摘要为了施行对转基因作物的监管,许多国家和机构制定了转基因标识制度的相关条例,因此建立针对转基因产品的检测方法是十分必要的。本研究运用环介导等温扩增技术(LAMP)建立了对转基因玉米(Zea mays L.) LY038外源基因的快速检测技术,该方法依据cordapA基因的序列设计了4条特异性引物,结果表明,引物特异性良好,该检测体系在63℃恒定温度下,反应50 min,可检测到0.01%的样本,是常规PCR方法的5倍。该检测方法具有高度的特异性、灵敏性和稳定性,该方法不需要特殊仪器、快速简便,在基层和现场检测中有很好的应用前景。
Abstract:Several countries and authorities have established relevant regulations on labeling for the surveillance of genetically modified organisms (GMOs). Therefore, it is imperative to establish appropriate methods to detect transgenic products. In this study, a loop-mediated isothermal amplification method (LAMP) was established for the rapid detection of genetically modified (GM) maize(Zea mays L.) LY038. Four LAMP primers were designed based on the cordapA gene. The LAMP assay was performed under isothermal conditions at 63℃ for 50 min. The specificity of the four primers was verified using eight other maize samples, in combination with enzymatic analysis using EcoR V to digest the LAMP product. All the results showed that the specificity of the primers was satisfying. To compare the detection limit of LAMP assay with PCR assay, seven kinds of different LY038 contents samples (100%, 10%, 1%, 0.5%, 0.1%, 0.05% and 0.01%) were prepared. The limit of detection of LAMP assay was 0.01%, which was 5-fold of traditional PCR assay. The established LAMP method has high specificity, high sensitivity and stability. Besides, it doesn’t need special equipment, and it is simple, which shows a good application prospect in primary and field use.