Molecular Characteristics and Epidemiological Significance of Shiga Toxin-Producing Escherichia coli O26 Strains

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Journal of Clinical Microbiology, June 2000, p. 2134-2140, Vol. 38, No. 6
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Molecular Characteristics and Epidemiological Significance of Shiga Toxin-Producing Escherichia coli O26 Strains

Wen-Lan Zhang,¹ Martina Bielaszewska,² Almut Liesegang,³ Helmut Tschäpe,³ Herbert Schmidt,¹ Martin Bitzan,⁴ and Helge Karch¹^,^*

Institut für Hygiene und Mikrobiologie der Universität Würzburg, 97080 Würzburg,¹ and Robert Koch Institut, Bereich Wernigerode, 38855 Wernigerode,³ Germany; Institute for Medical Microbiology, Charles University, Prague, 150 06 Prague, Czech Republic²; and Department of Pediatrics, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27157-1081⁴

Received 5 November 1999/Returned for modification 21 February 2000/Accepted 20 March 2000

Fifty-five Shiga toxin (Stx)-producing Escherichia coli (STEC) O26:H11 and O26:H strains isolated from humans between 1965 and 1999 in Germanyand the Czech Republic were investigated for their chromosomaland plasmid characteristics. All motile (n = 23) and nonmotile(n = 32) STEC O26 strains were shown to possess the identicalflagellin subunit-encoding gene (fliC). We observed a strikingrecent shift of the stx genotype from stx₁ to stx₂ among the STECO26 isolates. While stx₁ was the exclusive genotype identifiedin our collection until 1994, 94% of the isolates obtained after1997 possessed stx₂ either alone (71%) or together with stx₁ (23%).Plasmid profiling demonstrated a remarkable heterogeneity withrespect to plasmid sizes and combinations. Southern blot analysisof plasmid DNA with probes specific to potential accessory virulencegenes revealed considerable additional variability in gene compositionand arrangement. Pulsed-field gel electrophoresis (PFGE) differentiated16 subgroups among the 55 STEC O26 strains. Using these techniqueswe demonstrate the emergence of a new clonal subgroup characterizedby PFGE pattern A and a unique combination of virulence markersincluding stx₂ and a single, approximately 90-kb plasmid harboringthe enterhemorrhagic E. coli hlyA and etp genes. The proportionof PFGE subgroup A strains among STEC O26 isolates rose from 30%in 1996 to more than 50% in 1999. Four clusters of infectionswith the clonal subgroup A were identified. We conclude that theSTEC serogroup O26 is diverse and that pathogenic clonal subgroupscan rapidly emerge during short intervals. The extensive geneticdiversity of STEC O26 provides a basis for molecular subtypingof this important non-O157 STECserogroup.

^* Corresponding author. Mailing address: Institut für Hygiene und Mikrobiologie der Universität Würzburg, Josef-Schneider-Str.2, 97080 Würzburg, Germany. Phone: 49-931-201-5162. Fax: 49-931-201-5166.E-mail: hkarch{at}hygiene.uni-wuerzburg.de.

Journal of Clinical Microbiology, June 2000, p. 2134-2140, Vol. 38, No. 6
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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