Magnetic microbead enzyme-linked immunoassay based on phage encoded protein RBP 41-mediated for rapid and sensitive detection of Salmonella in food matrices

Rapid and sensitive quantitative detection methods are required to monitor and detect Salmonella throughout the food supply chain and early prevention of foodborne disease outbreaks. In this study, a magnetic microbead enzyme-linked immunoassay (MELISA) based on phage receptor binding protein was developed for rapid enrichment and detection of Salmonella in complex food matrices. RBP 41 from phage T102 acted as a species-specific recognition element for Salmonella by exploiting its strong binding capacity to Salmonella surface receptors. RBP 41-MBs were prepared by coupling recombinant RBP 41 with MBs and used to separate and enrich Salmonella cells from spiked food samples. The captured complexes were further integrated with ELISA procedures by HRP-labeled anti-Salmonella antibody for rapid and accurate detection of Salmonella. The whole method took <1.5 h and the detection limit was 10 CFU/mL. Therefore, MELISA was successfully developed for the detection of Salmonella in various spiked food samples (skim milk, lettuce, and chicken breast). The ELISA reaction process of this method was carried out on magnetic beads. It simplified the process of the traditional ELISA method and reduces the reaction time. This study expanded the use of phage-associated proteins and demonstrated the promising prospects for practical applications in the detection of foodborne pathogens.