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

This Small Business Innovation Research (SBIR) Phase II project proposes the development of novel enzymes (DNA polymerases) and other improvements for rapid detection of food-borne pathogens by DNA detection and amplification (PCR). PCR is a very fast and accurate method of pathogen detection, typically giving results in about a day, instead of several days required to grow and identify pathogens by cultural methods. But some foods, such as chocolate, dairy products, meat, and spices, contain components that inhibit the PCR assay. Current strategies for rapid pathogen testing in these foods include long cultural enrichment steps followed by dilution of inhibitors and/or labor intensive sample preparation (DNA extraction) to remove inhibitors. Inhibition-resistant DNA polymerases and food-specific PCR enhancers represent elegant, high-tech alternatives to dilution or DNA extraction. They could be integrated into existing rapid-detection systems to facilitate rapid accurate testing in inhibitory foods. The broader impacts of this research are reducing the number and severity of outbreaks of food-borne illnesses in the United States due to early detection of food-borne pathogens. Faster, more accurate detection of pathogens will save time and money for food manufacturers, and reduce the need for costly product recalls. Technology developed here could also extend the disciplines of forensics, where recovery of small amounts of DNA in the presence of a variety of inhibitors is critical, and national defense, where rapid detection of biological agents used as weapons could save lives.