Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 7/20/2004
Publication Date: 10/17/2004
Citation: Fratamico, P.M. 2004. Nucleic acid-based methods for detection and characterization of food-borne pathogens. River Falls Food Microbiology Symposium. Abstract. Paper # 14. Interpretive Summary:
Technical Abstract: Assays based on the polymerase chain reaction (PCR) are now accepted methods for rapidly confirming the presence or absence of specific pathogens in foods and other types of samples. Conventional PCR requires the use of agarose gel electrophoresis to detect the PCR product; whereas, real-time PCR combines DNA amplification with fluorescent probe detection of the amplified target sequence in a closed tube format. In our laboratory, single target and multiplex PCR assays, as well as real-time PCR assays have been developed for detection of E. coli O157:H7, Campylobacter species, simultaneous detection of E. coli O157:H7 and Salmonella species, Yersinia enterocolitica, and for specific detection of different E. coli serogroups based on unique gene sequences in the E. coli O antigen gene clusters. Microarrays, consisting of many probes complementary to pathogen-specific gene sequences bound to a solid substrate, can hybridize multiple DNA targets simultaneously; therefore, microarrays have tremendous potential for detection, identification, and characterization of pathogens. DNA arrays for typing of E. coli strains are currently in development. Novel methods combining on-chip PCR with simultaneous sequence-specific detection of amplification products on a solid phase show great potential for rapid, routine testing of bacterial pathogens in foods.