Presence of activatable Shiga toxin genotype (stx2d) in Shiga toxigenic Escherichia coli from livestock sources

Gobius, Kari S., Higgs, Glen M. and Desmarchelier, Patricia M. (2003) Presence of activatable Shiga toxin genotype (stx2d) in Shiga toxigenic Escherichia coli from livestock sources. Journal of Clinical Microbiology, 41 8: 3777-3783. doi:10.1128/JCM.41.8.3777-3783.2003

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Author Gobius, Kari S.
Higgs, Glen M.
Desmarchelier, Patricia M.
Title Presence of activatable Shiga toxin genotype (stx2d) in Shiga toxigenic Escherichia coli from livestock sources
Formatted title
Presence of activatable Shiga toxin genotype (stx2d) in Shiga toxigenic Escherichia coli from livestock sources
Journal name Journal of Clinical Microbiology   Check publisher's open access policy
ISSN 0095-1137
1098-660X
Publication date 2003
Sub-type Article (original research)
DOI 10.1128/JCM.41.8.3777-3783.2003
Open Access Status File (Publisher version)
Volume 41
Issue 8
Start page 3777
End page 3783
Total pages 7
Editor R. C. Tilton
Place of publication Washington, DC, United States
Publisher American Society for Microbiology
Collection year 2003
Language eng
Abstract Stx2d is a recently described Shiga toxin whose cytotoxicity is activated 10- to 1,000-fold by the elastase present in mouse or human intestinal mucus. We examined Shiga toxigenic Escherichia coli (STEC) strains isolated from food and livestock sources for the presence of activatable stx(2d). The stx(2) operons of STEC were first analyzed by PCR-restriction fragment length polymorphism (RFLP) analysis and categorized as stx(2), stx(2c) (vha), stx(2c) (vhb), or stx(2d) (EH250). Subsequently, the stx(2c) (vha) and stx(2c) (vhb) operons were screened for the absence of a PstI site in the stx(2a) subunit gene, a restriction site polymorphism which is a predictive indicator for the stx(2d) (activatable) genotype. Twelve STEC isolates carrying putative stx(2d) operons were identified, and nucleotide sequencing was used to confirm the identification of these operons as stx(2d). The complete nucleotide sequences of seven representative stx(2d) operons were determined. Shiga toxin expression in stx(2d) isolates was confirmed by immunoblotting. stx(2d) isolates were induced for the production of bacteriophages carrying stx. Two isolates were able to produce bacteriophages phi1662a and phi1720a carrying the stx(2d) operons. RFLP analysis of bacteriophage genomic DNA revealed that phi1662a and phi1720a were highly related to each other; however, the DNA sequences of these two stx(2d) operons were distinct. The STEC strains carrying these operons were isolated from retail ground beef. Surveillance for STEC strains expressing activatable stx(2d) Shiga toxin among clinical cases may indicate the significance of this toxin subtype to human health.
Keyword Microbiology
Hemolytic-uremic Syndrome
Variant B-subunit
Polymerase Chain-reaction
Cattle
Virulence
Strains
Genes
Prevalence
Cloning
Stec
Q-Index Code C1

 
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Created: Wed, 15 Aug 2007, 02:55:26 EST