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

Abstract

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.

Recombinase Polymerase Amplification for Diagnostic Applications

“…RPA was successfully used to detect major human pathogens including bacteria (6,(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23), viruses (9, 24 -32 ), fungi (33 ), and parasites (34,35 ), as well as genetically modified organisms (36,37 ) and genetic alterations observed in cancer cells (38,39 ). RPA was also used for HIV diagnosis in low-resource settings (40,41 ) (Tables 2 and 3).…”

Field Detection of Citrus Huanglongbing Associated with ‘Candidatus Liberibacter Asiaticus’ by Recombinese Polymerase Amplification within 15 min

“…(Figure S1). − Exponential amplification can be accomplished within 10 to 20 min by the circulation of this process. , RPA is a good choice for field detection because it is isothermal, simple, rapid, and sensitive. Application of RPA for detection of plant pathogens such as the Tomato mottle virus, Rose rosette virus , and Banana bunchy top virus have already been reported. − Russel and co-workers have reported implementing the rapid on-site detection of Las by RPA .…”