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Journal of Clinical Microbiology, May 2003, p. 2106-2112, Vol. 41, No. 5
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.5.2106-2112.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification and Characterization of a New Variant of Shiga Toxin 1 in Escherichia coli ONT:H19 of Bovine Origin

Institute for Hygiene and Technology of Food of Animal Origin, Veterinary Faculty, Ludwig-Maximilians-Universität, 80539 Munich,¹ Institute for Veterinary Food Science, Faculty of Veterinary Medicine, Justus-Liebig-Universität, 35392 Gießen, Germany²

Received 23 October 2002/ Returned for modification 27 December 2002/ Accepted 25 January 2003

A new variant of Shiga toxin 1 (Stx1), designated Stx1d, which deviates considerably more than any other known variant from Stx1 encoded by phage 933J, was identified in an Escherichia coli strain, ONT:H19, isolated from bovine feces. The complete stx₁ gene of this strain was amplified and sequenced. Nucleotide sequence homology with stx₁ from phage 933J was only 91%, resulting in the substitution of 20 amino acids in the A subunit and 7 amino acids in the B subunit of the protein. Cell culture supernatant of this strain, which was negative for stx₂ by PCR testing, was cytotoxic to Vero cells and gave positive results in two commercial enzyme-linked immunosorbent assays for Stx. PCR primers were constructed for the specific detection of the new variant. The findings of this study suggest that Stx1 is not as conserved as thought before and that there might be more variants which cannot be detected by commonly used PCR methods.

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