Development of Real-time PCR Methodology for the Rapid Enumeration of Low Numbers of Salmonella/Gram of Ground Beef Without Enrichment

The goal of these studies will be to develop methodology to allow the Rti-PCR reaction to detect low numbers (5 cells/g) of Salmonella in ground beef (fat content ranging 7 to 25 percent) within 4 hours without enrichment. Objectives will be to allow near real-time monitoring of ground beef with respect to the efficacy of sanitary practices and to rapidly obtain documentation to ensure that product shipped complies with new USDA standard for Salmonella in beef.
<P>
This project proposes to develop methodology for rapid PCR detection of low numbers of Salmonella in ground beef. The methodology to be developed will (1) effectively extract low numbers (5 CFU/g) of Salmonella in ground beef using b-cyclodextrin, and (2) will efficiently remove the extensive presence of PCR inhibitors by employing bentonite coated activated charcoal to remove such PCR inhibitors without binding the targeted bacterial cells in the aqueous extract.