A RAPID ASSAY FOR ESCHERICHIA COLI 0157:H7

THE PROBLEM ADDRESSED BY THIS PROPOSAL IS THE ACUTE NEED FOR A RAPID, SENSITIVE ASSAY FOR ESCHERICHIA COLI 0157:H7, AN EMERGING FOOD PATHOGEN CAPABLE OF CAUSING SEVERE ILLNESS, PERMANENT ORGAN DAMAGE, OR DEATH IN ITS VICTIMS. A PROGRAM FOR DEVELOPMENT OF AN ICE NUCLEATION GENE-TRANSDUCTION ASSAY FOR THIS ORGANISM IS DESCRIBED. SUCH AN ASSAY HAS ALREADY BEEN DEVELOPED FOR SALMONELLA, AND HAS BEEN SHOWN TO BE RAPID SENSITIVE, AND CONVENIENT. DURING PHASE I, THE SUITABILITY OF MULIPLE CANDIDATE PHAGES THAT INFECT E. COLI 0157:H7 FOR DEVELOPMENT INTO COMMERCIAL GENE-TRANSDUCING REAGENTS WILL BE EXAMINED. THE BEST PHAGE IDENTIFIED WILL THEN BE USED TO CONSTRUCT A DEMONSTRATION REAGENT THAT TRANSDUCES AN ICE NUCLEATION GENE INTO E. COLI 0157:H7. FINALLY, THE TARGET TRANSDUCTION EFFICIENCY AND CROSS-REACTIVITY TO NON-TARGET ORGANISMS OF THE DEMNSTRATION REAGENT WILL BE EVALUATED. SUCCESSFUL COMPLETION OF THE PROJECT WILL DEMONSTRATE THE FEASIBILITY OF PHASE II, WHERE THE DEMONSTRATION REAGENT WILL BE DEVELOPED INTO A HIGH-EFFICIENCY TRANSDUCING VEHICLE, AND THE RESULTING ASSAY WILL BE VALIDATED.