Location: Produce Safety and Microbiology Research
Project Number: 5325-32000-009-03
Start Date: Feb 02, 2012
End Date: Sep 30, 2014
The overall design includes three components: (1) cultural enrichment, (2) sandwich ELISA format assay using monoclonal antibodies, and (3) a microbead array platform. The improvements to the cultural enrichment step will be accomplished using incremental adjustments of existing standard microbiological media and do not concern this Agreement. An enrichment step is necessary to provide sufficient sensitivity for an assay that must detect the presence of only a few organisms in a sample of several hundred grams of food. The sandwich ELISA format we will employ is commonly performed in microplate format, and is typical for microbead immunochemical assays developed for the Luminex platform. We will use monoclonal and polyclonal antibodies provided by Romer Technologies. Each antigen assay will be deployed on a unique fluorescent labeled microbead set. Specificity and sensitivity will be confirmed using bacteria grown in culture broth. Validation will be performed by University of South Florida (USF) and FDA. For validation experiments pathogens will be spiked into foods, and the Lower Limit Of Detection will be determined for each organism, in the presence of other pathogens and typical background flora.