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Applied and Environmental Microbiology, March 2008, p. 1502-1507, Vol. 74, No. 5
0099-2240/08/$08.00+0     doi:10.1128/AEM.02125-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Detection of Viable but Nonculturable Escherichia coli O157:H7 Bacteria in Drinking Water and River Water

Division of Analytical and Environmental Toxicology, Department of Laboratory Medicine and Pathology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta T6G 2G3, Canada

Received 17 September 2007/ Accepted 3 January 2008

A sensitive method for specific detection of viable Escherichia coli O157:H7 cells, including viable but nonculturable (VBNC) cells, in water samples was developed. This method involved capture of the bacterial cells on a low-protein-binding membrane and direct extraction and purification of RNA followed by reverse transcription-PCR and electronic microarray detection of the rfbE and fliC genes of E. coli O157:H7. It detected as few as 1 CFU of E. coli O157:H7 in diluted cultures, 3 to 4 CFU/liter in tap water, 7 CFU/liter in river water, and 50 VBNC cells in 1 liter of river water, demonstrating the best limit of detection reported to date for VBNC cells in environmental water samples.

Published ahead of print on 18 January 2008.