Event-specific Real-time RPA Detection of Transgenic Rice Kefeng 6
1. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
2. Inspection and Testing Center for Environmental Risk Assessment of Genetically Modified Plant-related Microorganisms (Beijing), Ministry of Agriculture, Beijing 100081, China
GMO Biosafety Research, 2014, Vol. 5, No. 1 doi: 10.5376/gmo.2014.05.0001
Received: 07 Oct., 2014 Accepted: 30 Oct., 2014 Published: 12 Nov., 2014
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This article was first published in Fenzi Zhiwu Yuzhong, 12(5): 875-880 in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:
Xu et al., 2013, Event-specific Real-time RPA Detection of Transgenic Rice Kefeng 6, GMO Biosafety Research, Vol.4, No.1 1-5 (doi: 10.5376/gmo.2013.04.0001)
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.
GM crops detection; Isothermal amplification; RPA, Rice; Kefeng 6