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Journal of Clinical Microbiology, June 2005, p. 2944-2947, Vol. 43, No. 6
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.6.2944-2947.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Multiplex PCR for Direct Detection of Shiga Toxigenic Escherichia coli Strains Producing the Novel Subtilase Cytotoxin

Adrienne W. Paton* and James C. Paton

School of Molecular and Biomedical Science, University of Adelaide, Adelaide, South Australia 5005, Australia

Received 15 December 2004/ Returned for modification 4 February 2005/ Accepted 7 February 2005

We have recently described a novel AB5 subtilase cytotoxin produced by certain Shiga toxigenic Escherichia coli (STEC) strains. This potentially lethal toxin may contribute to severe gastrointestinal and systemic disease in humans. In this study we have developed a trivalent PCR assay for the detection of the novel toxin A subunit gene subA, as well as stx1 and stx2. The three primer pairs used in the assay do not interfere with each other and generate amplification products of 556, 180, and 255 bp, respectively. The assay can be used for determining the toxin genotype of STEC isolates, as well as for direct detection of toxin genes in primary fecal culture extracts.

* Corresponding author. Mailing address: School of Molecular and Biomedical Science, University of Adelaide, Adelaide, S.A. 5005, Australia. Phone: 61-8-83037552. Fax: 61-8-83033262. E-mail: adrienne.paton{at}adelaide.edu.au.

Journal of Clinical Microbiology, June 2005, p. 2944-2947, Vol. 43, No. 6
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.6.2944-2947.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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  • Hajra, T. K., Bag, P. K., Das, S. C., Mukherjee, S., Khan, A., Ramamurthy, T. (2007). Development of a Simple Latex Agglutination Assay for Detection of Shiga Toxin-Producing Escherichia coli (STEC) by Using Polyclonal Antibody against STEC. CVI 14: 600-604 [Abstract] [Full Text]  
  • Khaitan, A., Jandhyala, D. M., Thorpe, C. M., Ritchie, J. M., Paton, A. W. (2007). The Operon Encoding SubAB, a Novel Cytotoxin, Is Present in Shiga toxin-producing Escherichia coli Isolates from the United States. J. Clin. Microbiol. 45: 1374-1375 [Full Text]  
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