Abstract: This study combined enzymatic recombinase amplification (ERA) with lateral flow immunoassay (LFIA) to establish a rapid detection method for 11 high-risk pathogens in ready-to-eat foods, including Salmonella, Staphylococcus aureus, Escherichia coli O157:H7, Vibrio parahaemolyticus, Listeria monocytogenes, Shigella, Streptococcus, Pseudomonas aeruginosa, Yersinia enterocolitica, Cronobacter, and Burkholderia gladioli pv. cocovenenans. Primers and probes for each target pathogen were designed and screened, and the specificity, inclusivity, sensitivity, and applicability of the method to actual samples were evaluated. The results showed that the selected primers and probes for the 11 high-risk pathogens exhibited excellent specificity and inclusivity in the ERA-LFIA detection system. The limit of detection (LOD) for pure bacterial cultures ranged from 100 to 10-2 ng/μL. For artificially contaminated samples, the LOD was 1 CFU/mL following 4–8 h of enrichment. In commercially available ready-to-eat foods, the detection rates of Salmonella, V. parahaemolyticus, P. aeruginosa, and Y. enterocolitica were 16%, 24%, 8%, and 16%, respectively, which were consistent with the results obtained by the standard methods. The developed ERA-LFIA method is simple, rapid, accurate, and sensitive and offers the advantages of portability and visual readouts, which can provide technical support for the rapid screening and effective control of foodborne pathogens in ready-to-eat foods.

Key words: foodborne pathogens; enzymatic recombinase amplification; lateral flow immunoassay; rapid detection; ready-to-eat foods