Vedecký časopis - archív

48 2009

Journal of Food and Nutrition Research
Súhrny čísla 2 / 2009

Rapid and sensitive detection of pathogenic Yersinia enterocolitica strains in food using selective enrichment and real-time PCR
Journal of Food and Nutrition Research, 48, 2009, č. 2, s. 100-108

Eva Kaclíková, Department of Microbiology and Molecular Biology, VÚP Food Research Institute, P. O. Box 25, Priemyselná 4, SK – 82475 Bratislava 26, Slovakia.E-mail: kaclikova@vup.sk, tel.: +421-2-50237159, Fax: +421-2-55571417

Súhrn: Yersinia enterocolitica is an important human enteroinvasive pathogen with a global distribution. Food contamination is associated with pigs considered a major reservoir of Y. enterocolitica. The aim of this study was to develop and evaluate a rapid and sensitive real-time PCR-based method for the detection of pathogenic Y. enterocolitica in food. A TaqMan probe-based real-time PCR detection system targeting a sequence of chromosomally located ail (attachment and invasive locus) gene was designed. The real-time PCR system was evaluated as 100% selective for pathogenic Y. enterocolitica, which was determined using 20 Y. enterocolitica strains, 16 non-enterocolitica Yersinia spp. and 30 other Enterobacteriaceae strains. The PCR detection limit was 1  102 CFU.ml-1. Single selective enrichment based on the evaluation of various enrichment procedures was optimized. Out of 20 food samples analysed, four were positive by the PCR-based method and only one by the standard method. When 10 food matrices were artificially contaminated at a level of 100 CFU, ten and six sets of samples were positive by respective methods. The developed method may be used for the detection of pathogenic Y. enterocolitica as a specific and reliable alternative to the currently used microbiological method, furthermore providing a considerable reduction in the required analysis time.

Kľúčové slová: pathogenic Yersinia enterocolitica; real-time PCR; detection; food

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