Rapid detection of Escherichia coli O157:H7 in food using enrichment and real-time polymerase chain reaction
2010
Piskernik, S., University of Ljubljana, Ljubljana (Slovenia) | Klančnik, A., University of Ljubljana, Ljubljana (Slovenia) | Toplak, N., University of Ljubljana, Ljubljana (Slovenia) | Kovač, M., University of Ljubljana, Ljubljana (Slovenia) | Jeršek, B., University of Ljubljana, Ljubljana (Slovenia)
Escherichia coli O157:H7 may contaminate various types of meat products and cause diarrhea and vomiting, and also more serious complications such as haemorrhagic colitis and haemolytic-uremic syndrome (HUS) in humans. Traditional microbiological analyses to detect this pathogen are labour-intensive and time-consuming. The objective of this study was to evaluate a real-time polymerase chain reaction (PCR) for detection of E. coli O157:H7 in raw and ready-to-eat meat products. The detection limit of real-time PCR determined on pure culture was 1.1 x 10 sup(2) CFU per ml when DNA was obtained by lysing cells and 30.6 pgmicro per l when DNA was isolated and purified. Following a 20 h enrichment of a food sample in universal enrichment broth (UPB), the real-time PCR assay could detect 1.6 CFU per 10 g of E. coli O157:H7 in chicken juice, raw beef, minced beef, beefsteak tartare, brunch beef and beefburger with background flora in the range of less than10 sup(2)CFU per g to 2.1 x 10 sup(6) CFU per g. The applied method could be a useful tool for rapid detection of E. coli O157:H7 in raw meat and ready-to-eat meat products.
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