Submitted to: United States Japan Natural Resources Protein Panel
Publication Type: Proceedings
Publication Acceptance Date: September 17, 2004
Publication Date: October 17, 2004
Citation: Fratamico, P.M., Bagi, L.K., Liu, Y., Miyamoto, T. 2004. Nucleic acid-based methods for detection and analysis of foodborne pathogens. United States Japan Natural Resources Protein Panel. p. 348-353. Technical Abstract: Enterohemorrhagic E. coli (EHEC) serotype O157:H7 is a major cause of hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS) worldwide; however, numerous other Shiga toxin producing E. coli (STEC) serotypes have also caused similar illnesses and are found in similar foods and environmental niches as EHEC O157:H7. We have developed PCR-based assays, including multiplex PCR and real-time PCR assays targeting E. coli O157:H7 and other enterohemorrhagic E. coli. Furthermore, genetic-based detection and typing systems for E. coli strains are in development, involving sequencing of the E. coli O antigen gene clusters and design of serogroup-specific primers and probes for use in PCR- and DNA microarray-based assays. The sequence of the O antigen gene cluster of enterohemorrhagic E. coli O145 was determined, and multiplex PCR assays targeting the wzx and wzy genes found in the cluster and the stx1 and stx2 genes were developed and used to detect E. coli O145 in ground beef. The methods we are developing are useful for rapid detection, identification, and characterization of different E. coli strains and are needed to prevent the release of contaminated foods to the consumer, for epidemiological investigations, and to determine the prevalence of pathogenic E. coli in food, animals, and the environment.