荧光RPA技术检测转基因水稻科丰6号

Event-specific Real-time RPA Detection of Transgenic Rice Kefeng 6

目前转基因作物检测通常使用PCR检测方法,PCR检测耗时较长且需要精密控制温度循环的仪器,不适于现场检测。本研究应用重组酶聚合酶等温扩增技术（recombinase ploymerase amplification,RPA）,根据转基因抗虫水稻科丰6号插入连接区整合序列设计筛选了一套可用于RPA检测的引物及探针组合,建立了转基因抗虫水稻科丰6号及其衍生品种转化体特异性荧光RPA检测方法。研究结果表明本方法可稳定特异检出样品中500个拷贝的靶标分子,利用实时荧光检测简化了检测程序,使检测可在10~20 min内完成,与常规PCR检测需要数小时相比极大地缩短了检测时间。使用锂电池驱动的便携式扩增检测仪,对于野外转基因现场检测具有极大的应用价值。

GM crops detection commonly uses PCR method currently. It takes too long and needs thermal cycling instruments with accurate temperature control, so it is not suitable for field testing. In this study, event-specific detection method for transgenic insect resistance rice Kefeng 6 and its derivates based on real-time fluorescent recombinase ploymerase amplification （RPA） was established. According to the integration junction sequence of Kefeng 6, we designed and screened a set of RPA primers and probes combination which fit for RPA assays. The results showed that 500 copies of the target molecules in samples could be detected stablely and specifically by the method. The method can finish detection in 10 to 20 min, simplifying the test procedure by real-time fluorescence detection. Detection time was greatly shortened comparing with the conventional PCR detection which needed several hours. It has great application value for field transgenic detection using lithium battery-powered portable apparatus.