Monoclonal antibodies and an indirect ELISA for detection of psychrotrophic bacteria in refrigerated milk

Abstract

Monoclonal antibodies generated against live cells of Pseudomonas fluorescens have been used in an indirect ELISA format for the detection of Pseudomonas spp. and related psychrotrophic bacteria in refrigerated milk. The immunorecognition of monoclonal antibodies adsorbed to bacteria bound to the wells of a microtiter plate was performed with rabbit anti-mouse immunoglobulins conjugated to horseradish peroxidase. Subsequent enzymic conversion of the substrate resulted in distinct absorbance differences when assaying milk samples containing psychrotrophic bacteria in the range 105 to 109 CFU ml-1. The detection threshold for the ELISA assay developed in this work is 105 CFU ml-1.