Establishment of a Simple, Sensitive, and Specific Detection Method Based on Recombinase-Aided Amplification Combined with dsDNA-Specific Nucleases.

is a common foodborne pathogen that can cause food poisoning, posing a serious threat to human health. Therefore, quickly, sensitively, and accurately detecting is crucial to ensuring food safety. For the gene, we designed Recombinase-aided amplification (RAA) primers and dsDNA-specific nuclease (DNase) probes. The ideal primer and probe combination was found when conditions were optimized. Under UV light, a visual detection technique (RAA-dsDNase) was developed. Additionally, the RAA-dsDNase was modified to further reduce pollution hazards and simplify operations. One-pot RAA-dsDNase-UV or one-pot RAA-dsDNase-LFD was developed as a detection method, using UV or a lateral flow dipstick (LFD) for result observation. Among them, one-pot RAA-dsDNase and one-pot RAA-dsDNase-LFD had detection times of 50 min and 60 min, respectively, for detecting genomic DNA. One-pot RAA-dsDNase-UV had a detection limit of 10 copies/μL and 10 CFU/mL, while one-pot RAA-dsDNase-LFD had a sensitivity of 10 copies/μL and 10 CFU/mL. One-pot RAA-dsDNase-UV and one-pot RAA-dsDNase-LFD assays may identify 17 specific serovars witho ut causing a cross-reaction with the remaining 8 bacteria, which include . Furthermore, in tissue and milk samples has been reliably detected using both approaches. Overall, the detection method developed in this study can quickly, sensitively, and accurately detect , and it is expected to become an important detection tool for the prevention and control of in the future.