Rapid Detection of Mouse-Derived Components in Meat Products by Duplex Real-Time Recombinase-Aided Amplification

In this study, real-time recombinase-aided amplification (Rt-RAA) was used for the synchronous detection of rats-and mice-derived components in meat products. Primers and probes targeting the conserved sequences of the COX3 gene in rats and those of the 16S rRNA gene in mice were designed. The reaction parameters were optimized to establish the optimal reaction conditions for duplex Rt-RAA detection. Furthermore, the specificity, sensitivity and stability of the established duplex Rt-RAA method were evaluated, and it was applied to simulated commercial meat products. The results showed that this method had strong specificity and stability. The detection limits for rats- and mice-derived components were 0.5% and 0.2%, respectively. The detection limits for their synchronous detection were both 0.5%, and the detection limits for synchronous detection of rats- and mice-derived components mixed in simulated commercial meat samples were still as low as 0.5%. Using duplex Rt-RAA and real-time fluorescence quantitative polymerase chain reaction (real-time PCR), highly consistent results were obtained for commercially available samples and simulated samples. Therefore, the duplex Rt-RAA method is suitable for the detection of rats- and mice- derived components in meat products, providing a fast, efficient, and accurate new method for the detection of meat product adulteration.