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Journal of Clinical Microbiology, October 2003, p. 4671-4675, Vol. 41, No. 10
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.10.4671-4675.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Comparison of a Shiga Toxin Enzyme-Linked Immunosorbent Assay and Two Types of PCR for Detection of Shiga Toxin-Producing Escherichia coli in Human Stool Specimens

Matthias Pulz,1 Andreas Matussek,2,3 Masyar Monazahian,1 Andreas Tittel,1 Elisabet Nikolic,4 Maike Hartmann,5 Tobias Bellin,5 Jan Buer,3,5 and Florian Gunzer5*

Niedersächsisches Landesgesundheitsamt, 30449 Hannover,1 Cytonet GmbH & Co. Hannover Management KG,2 Institute of Medical Microbiology, Hannover Medical School, 30625 Hannover,5 Mucosal Immunity Group, German Research Centre for Biotechnology, 38124 Braunschweig, Germany,3 Department of Mathematics, University of Linköping, 581 83 Linköping, Sweden4

Received 8 November 2002/ Returned for modification 19 March 2003/ Accepted 10 June 2003

Shiga toxin (Stx)-producing Escherichia coli (STEC) is a major cause of sporadic cases of disease as well as serious outbreaks worldwide. The spectrum of illnesses includes mild nonbloody diarrhea, hemorrhagic colitis, and hemolytic-uremic syndrome. STEC produces one or more Stxs, which are subdivided into two major classes, Stx1 and Stx2. The ingestion of contaminated food or water, person-to-person spread, and contact with animals are the major transmission modes. The infective dose of STEC may be less than 100 organisms. Effective prevention of infection is dependent on rapid detection of the causative bacterial pathogen. In the present study, we examined 295 stool specimens for the presence of Stx-producing E. coli by three different methods: an Stx enzyme-linked immunosorbent assay, a conventional PCR assay, and a LightCycler PCR (LC-PCR) assay protocol recently developed by our laboratory at the Institute of Medical Microbiology at Hannover Medical School. Our intent was to compare these three methods and to examine the utility of the STEC LC-PCR protocol in a clinical laboratory. The addition of a control DNA to each sample to clearly discriminate inhibited specimens from negative ones enhanced the accuracy of the LC-PCR protocol. From our results, it can be concluded that LC-PCR is a very useful tool for the rapid and safe detection of STEC in clinical samples.

* Corresponding author. Mailing address: Institute of Medical Microbiology, Hannover Medical School, Carl-Neuberg Str. 1, 30625 Hannover, Germany. Phone: 49 511 532 4359. Fax: 49 511 532 4366. E-mail: fgunzer{at}mikbio.h.shuttle.de.

Journal of Clinical Microbiology, October 2003, p. 4671-4675, Vol. 41, No. 10
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.10.4671-4675.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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