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Yersinia enterocolitica is known to constitute a significant pathogen which possesses a huge impact on the pork production and processing industry in Poland. The Yersinia genus involves commensal microflora of pig intestines and may be easily transfer into carcasses during the improper and non hygienic slaughtering. Y. enterocolitica is considered to be responsible for provoking human diseases which includes self-limiting enteritis, acute mesenteric lymphadenitis, septicaemia, Reiter’s syndrome and erythema nodosum. These diseases mainly appear as the result of food-borne transmission. Pigs are believed to be the predominant reservoir of Y. enterocolitica strains which are pathogenic to humans. Furthermore, this pathogen can grow at low temperatures (even 0 °C) and survive at a wide pH range (5.0–9.6). Consumption of contaminated meat products constitutes the main route for Y. enterocolitica infections (Bolton et al., 2013). In Europe this pathogen is defined as the most common one provoking food-borne yersiniosis (Martinez, 2010). There is more non-pathogenic than pathogenic Yersinia strains identified in raw pork meat. Also, there is still a lack of reliable and rapid methods to identify  Y. enterocolitica since the conventional plating methods are not typically selective for Yersinia because they encourage the growth of other members of the family Enterobacteriaceae. Moreover, they are usually very laborious and time-consuming (Vázlerová and Steinhauserová, 2006; Van Damme et al., 2010). Real-time polymerase chain reaction (PCR) possesses the potential to cope with the significant limitations of conventional plating methods enabling the elimination of the pre-PCR enrichment step. 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 realtime 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. Received: 7 February 2017 Revised: 17 May 2017 Accepted: 8 June 2017

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