Detection of Escherichia coli O157:H7, Salmonella spp.,Staphylococcus aureus and Listeria monocytogenes in Kimchi byMultiplex Polymerase Chain Reaction (mPCR)

다국어 초록 (Multilingual Abstract)

We developed an mPCR assay for the simultaneous detection, in one tube, of Escherichia coli
O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes using species-specific
primers. The mPCR employed the E. coli O157:H7 specific primer Stx2A, Salmonella spp. specific
primer Its, S. aureus specific primer Cap8A-B and L. monocytogenes specific primer Hly.
Amplification with these primers produced products of 553, 312, 405 and 210 bp, respectively. All
PCR products were easily detected by agarose gel electrophoresis, and the sequences of the specific
amplicons assessed. Potential pathogenic bacteria, in laboratory-prepared and four commercially
available kimchi products, were using this mPCR assay, and the amplicons cloned and
sequenced. The results correlated exactly with sequences derived for amplicons obtained during
preliminry tests with known organisms. The sensitivity of the assay was determined for the purified
pathogen DNAs from four strains. The mPCR detected pathogen DNA at concentrations
ranging from approximately 0.45 to 0.05 pM/μl. Thus, this mPCR assay may allow for the rapid,
reliable and cost-effective identification of four potentially pathogens present in the mixed bacterial
communities of commercially available kimchi.

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