Graphene Oxide Nanosheet-Mediated Fluorescent RPA “Turn-On” Biosensor for rapid RNAi transgenic plant detection

Abstract In this paper preliminarily verified that graphene oxide (GO) nanomaterials enhanced the recombinase polymerase amplification (RPA). GO nanosheets improved the efficiency of RPA amplification by absorbing ingredients to induce local aggregation. The recombinase initially aggregated with the primers to form nucleoprotein filaments, absorbed on the GO nanosheets, changing the structure. Therefore, an isothermal fluorescence biosensor was developed based on GO nanosheets enhanced the RPA to detect RNA interference (RNAi) transgenic plants. FAM-labeled primers were absorbed and quenched by the GO nanosheets. After amplification, the primers were extended into double-stranded DNA, detaching from the GO surface to recover the fluorescent signal. The biosensor displayed high sensitivity and selectivity and showed an excellent relationship ranging from 1.5 to 100 ng of genome DNA, with a detection limit (LOD) of 1.5 ng. Consequently, the biosensor provides an enhanced isothermal method for detecting genetically modified (GM) products and exhibits significant potential for molecular detection.