VALIDATION OF RAPID TEST KITS FOR FOODBORNE PATHOGENS

The goal is to develop rapid test kits (RTKs) for E. coli (EC) O157:H7 and Salmonella Enteritidis (SE) via analytical verification and AOAC certification, for eventual commercialization of the RTKs. We have obtained antibody pairs for both targets from our preliminary screening studies. In this Phase I project, we conduct analytical verification of the RTKs built with these antibody pairs by:Determining the limit of detection (LOD) via spiking heat-killed bacteria culture into real food matrices.Determining specificity via challenging the RTKs with non-related to closely-related non-target bacteria (> 20 non-target organisms for each RTK).Conducting an accelerated stability test at 65°C to determine if the antibodies or enzyme conjugates are stable enough for 2 years storage at ambient temperature.Comparing with currently available commercial lateral flow kits, such as Neogen's Reveal 2.0 for EC O157:H7 and Romer Lab's RapidChek® SELECT™ Salmonella Enteritidis for SE.If any of these return unsatisfactory results, we may require further antibody screening, and/or optimization of enzyme-antibody conjugation method to improve sensitivity, specificity, and stability.