Cloning and Sequencing of a Shiga-like Toxin II-Related Gene from Escherichia coli O157: H7 Strain 7279

Meyer, Thomas; Karch, Helge; Hacker, Jörg; Bocklage, Hubertus; Heesemann, Jürgen

doi:10.1016/s0934-8840(11)80004-6

Cited by 40 publications

(39 citation statements)

References 30 publications

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“…1, lane 6). DNA sequence analysis of the stx 2 gene amplified from the Ont:H11 isolate showed 99.5% (5 nucleotide polymorphisms in the A subunit) sequence identity with stx 2vhb , a sequence variant of stx 2c (accession number X61283.1) (31). These data concur with the results of PCR-RFLP analyses used to type stx 2 variants in this study.…”

supporting

confidence: 88%

“…STEC can produce one or both of these groups of toxins which comprise two immunologically non-cross-reactive groups (designated Stx1 and Stx2), although other virulence factors such as intimin and enterohemolysin either directly contribute to (or are implicated by association with) the pathogenicity of STEC (11,21). stx 2 comprises 11 distinct variants (5,19,23,31,38,39,42,45,46,50) and is considered the most important STEC virulence factor associated with human disease (11,21,34,41). More importantly, differences in the degree of pathogenicity of STEC serotypes have been associated with variations in the stx 2 subtype (27,29,30,42).…”

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confidence: 99%

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Bovine Non-O157 Shiga Toxin 2-Containing Escherichia coli Isolates Commonly Possess stx _2-EDL933 and/or stx _2vhb Subtypes

et al. 2003

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stx 2 genes from 138 Shiga toxin-producing Escherichia coli (STEC) isolates, of which 127 were of bovine origin (58 serotypes) and 11 of human origin (one serotype; O113:H21), were subtyped. The bovine STEC isolates from Australian cattle carried ehxA and/or eaeA and predominantly possessed stx 2-EDL933 (103 of 127; 81.1%) either in combination with stx 2vhb (32 of 127; 25.2%) or on its own (52 of 127; 40.4%). Of 22 (90.9%) bovine isolates of serotype O113:H21, a serotype increasingly recovered from patients with hemolytic uremic syndrome (HUS) or hemorrhagic colitis, 20 contained both stx 2-EDL933 and stx 2vhb ; 2 isolates contained stx 2vhb only. Although 7 of 11 (63.6%) human O113:H21 isolates associated with diarrhea possessed stx 2-EDL933 , the remaining 4 isolates possessed a combination of stx 2-EDL933 and stx 2vhb . Three of the four were from separate sporadic cases of HUS, and one was from an unknown source.Shiga toxin-producing Escherichia coli (STEC) is commonly carried by ruminants (especially cattle) and is an important enteropathogen, causing human diseases ranging from mild diarrhea to more severe conditions such as hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS) (7, 15, 22a, 25, 28, 47). Although over 150 serotypes of STEC have been associated with human disease (12, 20), serotype O157:H7 is believed to be responsible for the majority of outbreaks and sporadic cases of HUS in the United States, Japan, and Europe. However, non-O157 STEC is of greater significance in Australia (16) and Argentina (35), and its role in HUS and HC in the United States and Europe is gaining recognition (24).The most critical factors of STEC associated with severe human disease are the Shiga toxins. STEC can produce one or both of these groups of toxins which comprise two immunologically non-cross-reactive groups (designated Stx1 and Stx2), although other virulence factors such as intimin and enterohemolysin either directly contribute to (or are implicated by association with) the pathogenicity of STEC (11,21). stx 2 comprises 11 distinct variants (5,19,23,31,38,39,42,45,46,50) and is considered the most important STEC virulence factor associated with human disease (11,21,34,41). More importantly, differences in the degree of pathogenicity of STEC serotypes have been associated with variations in the stx 2 subtype (27,29,30,42). The most frequently reported stx 2 subtypes are represented by stx 2c , stx 2d , and stx 2e (42,46,50).Although more than 200 different O:H serotypes of STEC have been isolated from cattle (1,7,12,20,24,49,51), there is a paucity of information regarding associations between serotype, stx 2 subtype, and virulence factor profiles among STEC isolates from meat-producing animals (9, 10, 22a, 26, 33, 43). stx 2 subtypes stx 2-EDL933 and stx 2c (stx 2vha and stx 2vhb ) have been described in European studies of bovine STEC; however, their association with serotype has not been reported previously (8,42,46). Ramachandran et al. (44) demonstrated that stx 2 -containing STEC from ovine f...

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Bovine Non-O157 Shiga Toxin 2-Containing Escherichia coli Isolates Commonly Possess stx _2-EDL933 and/or stx _2vhb Subtypes

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“…13 The Taq cycle sequencing method, which takes advantage of thermostable DNA polymerase, was found to be a potentially attractive method for precise genotyping of the genes belonging to the slt-11 family. However, if Taq-polymerase-mediated sequencing is performed before sequence analysis, digestion of the PCR products with FokI and HaeIII must be performed to exclude sequence errors due to the simultaneous presence of slt-II-and slt-II-related genes in the PCR products.…”

Section: Discussionmentioning

confidence: 99%

Genotyping of Shiga-like toxin genes in non-O157 Escherichia coli strains associated with haemolytic uraemic syndrome

Rüssmann

Kothe

Schmidt

et al. 1995

Journal of Medical Microbiology

103

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Summary.The pheno-and genotypes of Shiga-like toxins (SLTs) in non-0157 Escherichia coli strains from patients with haemolytic uraemic syndrome were determined. The clinical isolates investigated were from Italy and Germany and belonged to serotypes 022: H8, 026 : H-, 026 : HI 1, 091 : H-, 0 1 1 1 : H-and 0128 : H-; one isolate was non-typable. SLT genotypes were analysed by complete nucleotide sequence analysis of the B-subunit genes. The results showed that 14 strains possessed slt-I alone, two contained slt-II alone and five isolates harboured both slt-I and slt-ZI genes. In only two strains were slt-11-related genes found, together with either slt-Z or slt-II. These findings indicate that variants of SLT-I1 are rarely found in non-0157 E. coli isolates from patients with haemolytic uraemic syndrome. Polymerase chain reaction (PCR) with Taq cycle sequencing was found to be a suitable method for classification of slt genotypes.

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“…The sequences of the stx 2 gene from two phages were more similar to 933W, whereas the gene from five phages showed higher identity with a gene encoding Shiga-like toxin type IIvhc, accession number X61283 (Meyer et al, 1992), currently known as stx 2c (WHO, 1991). The remainder showed very high identity with the gene sequence of phage Ni12 (El-Sayed, 2002), induced from an E. coli strain isolated from a bovine source.…”

Section: Sequencing Of the Stx 2 Gene Encoded By Temperate Phagesmentioning

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Diversity of stx 2 converting bacteriophages induced from Shiga-toxin-producing Escherichia coli strains isolated from cattle

et al. 2004

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The presence of bacteriophages encoding Shiga toxin 2 (stx 2 phages) was analysed in 168 strains of Shiga-toxin-producing Escherichia coli (STEC) isolated from cattle. Following mitomycin C induction, strains carrying stx 2 phages were screened by plaque blot and hybridization with an stx 2 A-probe. In the stx 2 -phage-carrying strains, the amounts of phage production, phage DNA extracted and Stx 2 produced after induction were assessed. The induced stx 2 phages were characterized morphologically and genetically. Assays to obtain lysogens from different strains were also carried out and phages induced from the lysogens were compared with those induced from the STEC isolates. Results indicated that 18 % of the strains carried an inducible stx 2 phage. Most of them showed a direct relationship between phage induction and toxin production. Each strain carried only one inducible stx 2 phage, although a few strains had two copies of the stx 2 in the chromosome. The stx 2 phages showed diverse morphology and a wide variability in their genome. Assays to obtain lysogens showed that not all the phages were transduced with the same frequency and only six lysogens were obtained. Phages in the lysogens were the same as those induced from their respective initial STEC host strains, although the induction and relative toxin production of the lysogens varied. Most phages carried the stx 2 gene, while a few carried stx 2 variants. Infectivity of the phages depended on the different hosts, although O157 : H7 was preferentially infected by phages induced from O157 strains. The results show that inducible stx 2 phages are common among STEC of animal origin and that they may enhance the spread of stx 2 .

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Cloning and Sequencing of a Shiga-like Toxin II-Related Gene from Escherichia coli O157: H7 Strain 7279

Cited by 40 publications

References 30 publications

Bovine Non-O157 Shiga Toxin 2-Containing Escherichia coli Isolates Commonly Possess stx _2-EDL933 and/or stx _2vhb Subtypes

Bovine Non-O157 Shiga Toxin 2-Containing Escherichia coli Isolates Commonly Possess stx _2-EDL933 and/or stx _2vhb Subtypes

Genotyping of Shiga-like toxin genes in non-O157 Escherichia coli strains associated with haemolytic uraemic syndrome

Diversity of stx 2 converting bacteriophages induced from Shiga-toxin-producing Escherichia coli strains isolated from cattle

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Cloning and Sequencing of a Shiga-like Toxin II-Related Gene from Escherichia coli O157: H7 Strain 7279

Cited by 40 publications

References 30 publications

Bovine Non-O157 Shiga Toxin 2-Containing Escherichia coli Isolates Commonly Possess stx 2-EDL933 and/or stx 2vhb Subtypes

Bovine Non-O157 Shiga Toxin 2-Containing Escherichia coli Isolates Commonly Possess stx 2-EDL933 and/or stx 2vhb Subtypes

Genotyping of Shiga-like toxin genes in non-O157 Escherichia coli strains associated with haemolytic uraemic syndrome

Diversity of stx 2 converting bacteriophages induced from Shiga-toxin-producing Escherichia coli strains isolated from cattle

Contact Info

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Bovine Non-O157 Shiga Toxin 2-Containing Escherichia coli Isolates Commonly Possess stx _2-EDL933 and/or stx _2vhb Subtypes

Bovine Non-O157 Shiga Toxin 2-Containing Escherichia coli Isolates Commonly Possess stx _2-EDL933 and/or stx _2vhb Subtypes