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United States Department of Agriculture

Agricultural Research Service

Title: Rapid Pcr Detection of Enterohemorrhagic Escherichia Coli (Ehec) in Bovine Food Products and Feces

Authors
item Ellingson, Jay - MARSHFIELD CLINIC LABS
item Koziczkowski, Jeff - MARSHFIELD CLINIC LABS
item Anderson, Jennifer - MARSHFIELD CLINIC LABS
item Carlson, Steven
item Sharma, Vijay

Submitted to: Molecular and Cellular Probes
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 12, 2005
Publication Date: June 20, 2005
Citation: Ellingson, J.L.E., Koziczkowski, J.J., Anderson, J.L., Carlson, S.A., Sharma, V.K. 2005. Rapid PCR detection of enterohemorrhagic Escherichia coli (EHEC) in bovine food products and feces. Molecular and Cellular Probes. 19(3):213-217.

Interpretive Summary: Although Escherichia coli (E. coli) O157:H7 is a major cause of foodborne illness, other types of E. coli can also cause illness. Many genetic assays have been developed to detect Enterohemorrhagic E. coli (EHEC), including E. coli O157:H7. However, no assay currently exists to detect E. coli O157:H7 and other EHEC strains with a confirmed positive or negative result in less than 12 hours. Raw beef food products (raw ground beef and raw boneless beef) and bovine fecal samples were inoculated with E. coli O157:H7 to test the sensitivity of this assay. Fourteen different foodborne bacteria, including E. coli O157:H7, and 19 varrious E. coli strains, obtained from the United States Department of Agriculture, Agricultural Research Service (USDA-ARS) were tested for specificity. E. coli O157:H7 was detected at very low levels in less than 12 hours. None of the 14 foodborne bacteria screened for cross-reactivity was detected.

Technical Abstract: Although Escherichia coli (E. coli) O157:H7 is a major cause of foodborne illness, other types of E. coli can also cause illness. E. coli that possess the eae gene for attachment and effacing have the potential to cause disease. Many real-time, molecular-based assays have been developed to detect Enterohemorrhagic E. coli (EHEC), including E. coli O157:H7. However, no assay currently exists to detect the eae gene present in E. coli O157:H7 and other EHEC strains with a confirmed positive or negative result in less than 12 hours. Raw beef food products (raw ground beef and raw boneless beef) at 25 g and 375 g samples and bovine fecal samples at 2 g were inoculated with 10**1, 10**3, 10**4, and 10**5 organisms of E. coli O157:H7 to test the sensitivity of this assay. Fourteen different foodborne bacteria (including E. coli O157:H7) and nineteen various E. coli strains, obtained from the United States Department of Agriculture, Agricultural Research Service (USDA-ARS) were tested for specificity. E. coli O157:H7 was detected at the level of 10**1 organisms in both 25 g and 375 g samples of raw ground and raw boneless beef products as well as 2 g samples of bovine feces after pre-enrichment and concentration. None of the fourteen foodborne bacteria screened for cross-reactivity was detected. All USDA E. coli strains confirmed to contain the eae gene were detected.

Last Modified: 7/28/2014
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