Simultaneous detection of six food-borne pathogens by multiplex PCR with a GeXP analyzer

Abstract A novel application of GeXP analyzer was developed for simultaneous detection of six pathogens associated with food poisoning outbreaks, including Salmonella enterica, Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, Shigella spp., and Campylobacter jejuni . Chimeric primers containing both microbe- and pMD19-specific sequences were fused to universal sequences resulted in PCR products with intended sizes. The PCR products were separated by capillary electrophoresis and identified by using fluorescence spectrophotometry. Plasmid pMD19-T was added to each reaction as positive control to ascertain the PCR steps and data analysis step of the assay. The results indicate that the GeXP method is both specific and sensitive for the detection of all six food-borne pathogens in a single reaction-without any enrichment step. In conclusion, the GeXP-PCR assay is a rapid, sensitive and high-throughput method for parallel analysis of food-borne pathogens.