Particle concentration flourescence immunoassay (PCFIA) for rapid detection of Salmonella
1995
Garcia, A.V. | Brooks, B. (Animal Diseases Research Inst., Nepean, Ontario (Canada). Food Production and Inspection Branch)
A two antibody sandwich PCFIA was developed for the rapid detection of Salmonella in chicken carcass rinses. The PCFIA system utilized sub-micron polystyrene beads coated with rabbit anti-Salmonella LPS O-chain polyclonal antibodies to capture Salmonella antigen. A murine monoclonal antibody (M178) specific for Salmonella core LPS followed by an FITC conjugated rabbit anti-mouse polyclonal antiserum was used for detection. Studies were conducted to evaluate capture and detection antibody concentrations, antigen treatment conditions including heating in the presence of proteinase K or sodium deoxycholate, and the use of sonication to obtain a homogenous suspension of the polysterene beads during the reaction and washing steps. Representatives of four Salmonella serogroups were used to determine the detection level of the assay. With pure cultures of S. typhimurium (Group B), S. thompson (Group C), S. enteritidis (Group D), and S. anatu (Group E) the detection limit was 1 x 10 raised to the seventh power CFU/mL. The limit of detection in chicken carcass rinses spiked with S. thompson and S. anatum was 1 x 10 raised to the eight power CFU/mL and 5 x 10 raised to the eight power CFU/mL in rinses spiked with S. typhimurium and S. enteritidis. The monoclonal antibody used in the PCFIA reacted with all Salmonella spp. tested and gave only minimal background levels with carcass rinse samples in which no Salmonella were detected using the standard culture procedures. The results indicate that the PCFIA is efficient to detect Salmonella in chicken carcass rinses
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