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The aim of this study was to compare a real-time PCR assay, with a conventional culture/PCR method, to detect  S. aureus ,  mecA  and Panton-Valentine Leukocidin (PVL) genes in animals and retail meat, using a two-step selective enrichment protocol. A total of 234 samples were examined (77 animal nasal swabs, 112 retail raw meat, and 45 deli meat). The multiplex real-time PCR targeted the genes:  nuc  (identification of  S. aureus ),  mecA  (associated with methicillin resistance) and PVL (virulence factor), and the primary and secondary enrichment samples were assessed. The conventional culture/PCR method included the two-step selective enrichment, selective plating, biochemical testing, and multiplex PCR for confirmation. The conventional culture/PCR method recovered 95/234 positive  S. aureus  samples. Application of real-time PCR on samples following primary and secondary enrichment detected  S. aureus  in 111/234 and 120/234 samples respectively. For detection of  S. aureus , the  kappa  statistic was 0.68–0.88 (from substantial to almost perfect agreement) and 0.29–0.77 (from fair to substantial agreement) for primary and secondary enrichments, using real-time PCR. For detection of  mecA  gene, the  kappa  statistic was 0–0.49 (from no agreement beyond that expected by chance to moderate agreement) for primary and secondary enrichment samples. Two pork samples were  mecA  gene positive by all methods. The real-time PCR assay detected the  mecA  gene in samples that were negative for  S. aureus , but positive for  Staphylococcus  spp. The PVL gene was not detected in any sample by the conventional culture/PCR method or the real-time PCR assay. Among  S. aureus  isolated by conventional culture/PCR method, the sequence type ST398, and multi-drug resistant strains were found in animals and raw meat samples. The real-time PCR assay may be recommended as a rapid method for detection of  S. aureus  and the  mecA  gene, with further confirmation of methicillin-resistant  S. aureus  (MRSA) using the standard culture method.

plos one

Multiplex Real-Time PCR for Detection of Staphylococcus aureus, mecA and Panton-Valentine Leukocidin (PVL) Genes from Selective Enrichments from Animals and Retail Meat