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http://dx.doi.org/10.5851/kosfa.2014.34.5.717

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

Kim, Ji-Hyun (Department of Food Science and Biotechnology of Animal Resources, Konkuk University)
Rhim, Seong-Ryul (Department of Food Science and Biotechnology of Animal Resources, Konkuk University)
Kim, Kee-Tae (Bio/Molecular Informatics Center, Konkuk University)
Paik, Hyun-Dong (Department of Food Science and Biotechnology of Animal Resources, Konkuk University)
Lee, Joo-Yeon (Korea Livestock Products HACCP Accreditation Service)
Publication Information
Food Science of Animal Resources / v.34, no.5, 2014 , pp. 717-723 More about this Journal
Abstract
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 to increase sensitivity. Multiplex PCR developed in this study produced only actA, fliC, hbl, invA, ileS amplicons, but did not produce any non-specific amplicon. The primer sets successfully amplified the target genes in the multiplex PCR without any non-specific or additional bands on the other strains. The multiplex PCR assays also amplified some target genes from five pathogens, and multiplex amplification was obtained from as little as 1 pg of DNA. According to the results from the sensitivity evaluation, the multiplex PCR developed in this study detected 10 cells/mL of the pathogens inoculated in milk samples, respectively. The results suggested that multiplex PCR was an effective assay demonstrating high specificity for the simultaneous detection of five target pathogens in food system.
Keywords
simultaneous detection; multiplex PCR; pathogens; low-fatted milk;
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