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The immediate goal of this seed grant proposal is to determine the best target site for a model pathogenic organism and assess the value of PNA-based assay systems for this application. The long-term goal of this project is to develop a panel of microchip-based "lab-on-a-chip" assays with immediate (in minutes) results that would also be simple, user-friendly and low-cost to facilitate onsite testing.
Loss of consumer confidence stemming from highly publicized deaths and illnesses from Vibrio vulnificus and V.parahaemolyticus has caused a decline in the shellfish industry. Monitoring and mitigation steps taken at the primary producer level, prior to secondary treatment or marketing, could increase profitability and reduce product recalls. Therefore there is a need for improved methods capable of rapid, onsite analysis. Detection and enumeration of Vibrios is currently limited to extensive microbiological procedures or complex molecular analysis requiring skill and expensive equipment. The model organism will be V. parahaemolyticus, a pathogen that has caused increased human illness and mortalities in the US and worldwide over the last decade. This project represents an effort to transfer newly emerging rapid detection technology to food, environmental and clinical samples in order to develop rapid, robust, specific, sensitive and user-friendly protocols (the faster, cheaper, better model). These biodetectors are based on the properties of the synthetic DNA analog peptide nucleic acid (PNA). PNA's have unique binding properties of more rapid, specific and tighter binding to target DNA.