Simultaneous Detection of Listeria monocytogenes, Escherichia coli O157:H7, Bacillus cereus, Salmonella spp., and Staphylococcus aureus in Low-fatted Milk by Multiplex PCR

A rapid and specific PCR assay for the simultaneous detection of Listeria monocytogenes, Escherichia coli O157:H7,Bacillus cereus, Salmonella spp., and Staphylococcus aureus in foods was developed to reduce the detection time and toincrease sensitivity. Multiplex PCR developed in this study produced only actA, fliC, hbl, invA, ileS amplicons, but did notproduce any non-specific amplicon. The primer sets successfully amplified the target genes in the multiplex PCR withoutany non-specific or additional bands on the other strains. The multiplex PCR assays also amplified some target genes fromfive pathogens, and multiplex amplification was obtained from as little as 1 pg of DNA. According to the results from thesensitivity evaluation, the multiplex PCR developed in this study detected 10 cells/mL of the pathogens inoculated in milksamples, respectively. The results suggested that multiplex PCR was an effective assay demonstrating high specificity forthe simultaneous detection of five target pathogens in food system.