Development of an Indirect Competitive Enzyme-linked Immunosorbent Assays Method Based on Immunomagetic-bead for Analyzing Listeria monocytogenes in Food

摘要

Listeria monocytogenes is an emerging bacterial food borne pathogen, which could survive common stress levels and often cause listeriosis. To control its infection, it is necessary to establish a sensitive, specific, rapid technology for detecting Listeria monocytogenes . In this study, invasion-associated-protein (IAP) was used as a target gene. After prokaryotic cloning and expression of the gene, the specific membrane protein P60 of Listeria moncytogenes was obtained. The P60 was injected into New Zealand white rabbits to get the antibody. The prepared antibody was conjugated with magnetic-bead to get immunomagnetic-beads. Enzyme-linked immunosorbent assays method based on immunomagetic-bead (IMB-ELISA) would be developed through immunomagetic-bead (IMB) collecting Listeria moncytogenes . The results showed that the sensitivity of the IMB-ELISA was 1.2x10⁶ CFU mL^?1, the limit of detection was 31 CFU mL^?1. The IMB-ELISA is a quick and accurate method for the detection of Listeria moncytogenes .