Quantitative assessment of Yersinia enterocolitica in raw pork meat using real time PCR (qPCR) technique

Abstract

The aim of the present study was to develop a highly sensitive and specific TaqMan probe with a set of primers for detection of pathogenic Yersinia enterocolitica strains and to eliminate the pre-PCR enrichment step in the real-time PCR (qPCR). A newly developed qPCR assay which is sensitive and specific for quick and reliable quantitative assessment of Y. enterocolitica present in artificially contaminated raw pork meat samples is described. This protocol involves the qPCR method with a TaqMan probe. The primers and probe were designed on the base of locus-tag CH49-3099 gene. This protocol appeared to be reliable for both intended applications: 1. identification and quantification of Y. enterocolitica in artificially and naturally contaminated raw pork meat and 2. establishment of growth potentials of different serotypes of Y. enterocolitica in raw meat at the usually used storage temperatures. This developed method makes it possible to eliminate the pre-PCR enrichment step enabling for the rapid assessment of meat-related consumer exposure to this pathogen.