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Applied and Environmental Microbiology, February 1999, p. 868-872, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Detection of Shiga-Like Toxin (stx1 and stx2), Intimin (eaeA), and Enterohemorrhagic Escherichia coli (EHEC) Hemolysin (EHEC hlyA) Genes in Animal Feces by Multiplex PCR

Peter K. Fagan,1 Michael A. Hornitzky,1 Karl A. Bettelheim,2 and Steven P. Djordjevic1,*

Elizabeth Macarthur Agricultural Institute, New South Wales Agriculture, Camden, New South Wales 2570,1 and Victorian Infectious Diseases Reference Laboratory, Fairfield Hospital, Fairfield, Victoria 3078,2 Australia

Received 8 June 1998/Accepted 20 November 1998

A multiplex PCR was developed for the rapid detection of genes encoding Shiga toxins 1 and 2 (stx1 and stx2), intimin (eaeA), and enterohemolysin A (hlyA) in 444 fecal samples derived from healthy and clinically affected cattle, sheep, pigs, and goats. The method involved non-solvent-based extraction of nucleic acid from an aliquot of an overnight culture of feces in EC (modified) broth. The detection limit of the assay for both fecal samples and pure cultures was between 18 and 37 genome equivalents. stx1 and hlyA were the most commonly encountered virulence factors.

* Corresponding author. Mailing address: Elizabeth Macarthur Agricultural Institute, New South Wales Agriculture, Private Mail Bag 8, Camden, New South Wales 2570, Australia. Phone: 0061-246-406426. Fax: 0061-246-406384. E-mail: steve.djordjevic{at}agric.nsw.gov.au.

Applied and Environmental Microbiology, February 1999, p. 868-872, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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