The objective of this study was to develop a multiplex real-time polymerase chain reaction (PCR) melt assay to detect and identify three food-borne pathogens Salmonella enterica, Escherichia coli and Shigella flexneri without post-PCR processing such as gel or capillary electrophoresis. Phylogenetic analyses of these three genera have proven that their genomes are very similar. In fact, the genomes of many Shigella strains are ancestral clones of Escherichia strains. Primers were generated targeting the origin of replication, virA gene, and lacY gene for Salmonella sp., S. flexneri, and E. coli, respectively, provide unique amplicons for differential analysis. ATCC reference strains were used to evaluate the assay. Specific amplicons were obtained from all species.

Multiplex real-time PCR detection and identification of food-borne pathogens Salmonella enterica, Escherichia coli and Shigella flexneri

249th National Meeting (2015)

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