SBIR Phase I: Inhibition-Resistant DNA Polymerases and Other Improvements for Detection of Food-borne Pathogens.

This Small Business Innovation Research (SBIR) Phase I project proposes the improvement of real-time PCR, a DNA-based rapid-detection method, for detection of food-borne pathogens such as Salmonella. Current methods are inadequate to accurately detect pathogens in foods such as chocolate, soft-cheese, and milk, which are inhibitory to PCR. These foods present a challenge due to their interference with the sensitivity of the assay. Longer, more labor-intensive tests are required for these foods, as inaccurate detection can lead to potentially deadly false negatives. The proposed improvements will shorten and simplify pathogen detection for these foods. The novel inhibition-resistant mutant enzymes and enhancers can to applied to PCR, and simplify other stages of current protocols such as cultural enrichment (an incubation period usually required for pathogen detection) and sample preparation. These improvements may also allow for more rapid, accurate testing for a broad range of foods, not only those foods known to be inhibitory to PCR. The broader impacts of this research include a safer food supply, due to the faster, more accurate detection of food-borne pathogens. This will allow food producers and manufacturers to more quickly respond to any discovered contamination, and facilitate immediate recalls of infected food. For routine tests, these innovations will give testing labs accurate and reliable results, while saving them time, labor, and money.