A rapid (one day), sensitive real-time polymerase chain reaction assay for detecting Escherichia coli O157:H7 in ground beef |
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Authors: | Holicka Jane Guy Rebecca A Kapoor Anita Shepherd David Horgen Paul A |
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Institution: | Master of Biotechnology Program, Department of Biology, University of Toronto at Mississauga, ON, Canada. |
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Abstract: | The purpose of this study was to apply our rapid, integrated double enrichment 5' nuclease real-time polymerase chain reaction assay for the detection of Escherichia coli O157:H7 and evaluate its efficacy. The assay targeted ground beef, an important vehicle in disease epidemiology. The assay reliably determined in 8 h the presence of E. coli O157:H7 in ground beef at the level of 1 colony-forming unit (CFU)/g. The sensitivity and specificity of the assay were compared with that of standard enrichment diagnostic techniques. A correlation of 100% in detection was achieved to the limit of 1 CFU/g. This assay can be used as a rapid, automatic process for identification of E. coli O157:H7 in ground beef or can be integrated with standard culture procedures, resulting in considerable cost and time savings. |
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