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Journal of Clinical Microbiology, November 2001, p. 3992-3998, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.3992-3998.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Isolation of a Lysogenic Bacteriophage Carrying the stx1OX3 Gene, Which Is Closely Associated with Shiga Toxin-Producing Escherichia coli Strains from Sheep and Humans

Claudia Koch,1 Stefan Hertwig,1 Rudi Lurz,2 Bernd Appel,1 and Lothar Beutin1,*

Robert Koch-Institut, D-13353 Berlin,1 and Max-Planck-Institut für Molekulare Genetik, D-14195 Berlin,2 Germany

Received 12 April 2001/Returned for modification 4 July 2001/Accepted 21 August 2001

A specific PCR for the detection of a variant of the gene encoding Shiga toxin 1 (stx1) called stx1OX3 (GenBank accession no. Z36901) was developed. The PCR was used to investigate 148 Stx1-producing Escherichia coli strains from human patients (n = 72), cattle (n = 27), sheep (n = 48), and a goat (n = 1) for the presence of the stx1OX3 gene. The stx1OX3 gene was present in 38 Shiga toxin-producing E. coli (STEC) strains from sheep belonging to serogroups O5, O125, O128, O146, and OX3 but was absent from Stx1-positive ovine STEC O91 strains. The stx1OX3 gene was also detected in 22 STEC strains from humans with nonbloody diarrhea and from asymptomatic excreters. Serotypes O146:H21 and O128:H2 were most frequently associated with stx1OX3-carrying STEC from sheep and humans. In contrast, Stx1-producing STEC strains from cattle and goats and 50 STEC strains from humans were all negative for the stx1OX3 gene. The stx1OX3-negative strains belonged to 13 serotypes which were different from those of the stx1OX3-positive STEC strains. Moreover, the stx1OX3 gene was not associated with STEC belonging to enterohemorrhagic E. coli (EHEC) serogroups O26, O103, O111, O118, O145, and O157. A bacteriophage carrying the stx1OX3 gene (phage 6220) was isolated from a human STEC O146:H21 strain. The phage was able to lysogenize laboratory E. coli K-12 strain C600. Phage 6220 shared a similar morphology and a high degree of DNA homology with Stx2-encoding phage 933W, which originates from EHEC O157. In contrast, few similarities were found between phage 6220 and Stx1-encoding bacteriophage H-19B from EHEC O26.

* Corresponding author. Mailing address: (P13) Division of Emerging Bacterial Pathogens, Robert Koch-Institut, Nordufer 20, D-13353 Berlin, Germany. Phone: 49 30 45 47 2484. Fax: 49 30 45 47 2673. E-mail: BeutinL{at}rki.de.

Journal of Clinical Microbiology, November 2001, p. 3992-3998, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.3992-3998.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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