Rapid Detection of Listeria monocytogenes Using a Fluorescence Immunochromatographic Assay Combined with Immunomagnetic Bead Separation
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Abstract:
In order to establish a simple, rapid, sensitive, and specific method to detect Listeria monocytogenes, in this study, immunomagnetic beads were prepared by coupling anti-L. monocytogenes monoclonal antibody with magnetic beads. The fluorescence immunochromatographic strips were composed of anti-L. monocytogenes polyclonal antibody and mouse IgG marked by fluorescent microspheres as the detection antibody, anti-L. monocytogenes polyclonal antibody as the test line, and the goat anti-mouse IgG secondary antibody as the control line. Immunomagnetic separation was combined with a fluorescence immunochromatographic assay, and this method was applied to rapidly detect L. monocytogenes. The results showed that the detection limit of fluorescence immunochromatographic strips for pure cultures was 4 × 105 CFU/mL. For the joint detection method using samples concentrated 10- and 100-fold, the detection limits of pure culture samples were 4 × 104 CFU/mL and 1 × 104 CFU/mL, respectively. The joint detection method showed good specificity, and no cross-reactivity of the 10 strains kept in the laboratory was observed. The detection limit of artificially contaminated samples was also 1 × 104 CFU/mL and was not reduced compared with pure cultures. This method is of great value for rapid on-site detection of L. monocytogenes in food products.
