title: Variants of Shiga-like toxin II constitute a major toxin component in Escherichia coli O157 strains from patients with haemolytic uraemic syndrome.
abstract: The prevalence and genotype of Shiga-like toxins (SLTs) in Escherichia coli (O)157 strains from patients in Germany with haemolytic uraemic syndrome (HUS) were investigated. This was done by PCR amplification of the B-subunit genes with two primer pairs--one complementary to slt-IB, and the other homologous to both slt-IIB and slt-IIvB sequences. To distinguish between slt-II and slt-IIv, the amplified DNA was digested with restriction endonucleases HaeIII and FokI. Of the 38 strains examined, 17 harboured sequences for slt-IIv; four contained only slt-IIv, three carried both slt-IIv and slt-I, and 10 strains had slt-IIv and slt-II. A further three genotypes (slt-I, slt-II, slt-I/slt-II) were found in the remaining 21 strains resulting in a total of six slt genotypes. To determine whether the slt genes were expressed, and whether genotypes correlated with phenotypes, all strains were subjected to cytotoxicity assays and colony ELISA. All 38 strains displayed cytotoxic activity to Vero cells in similar quantities. The SLT-I-specific monoclonal antibody (MAb)13C4 reacted with all 10 strains in which slt-I sequences were identified. Colony blot ELISA with the SLT-II specific MAb11E10 detected 27 of 28 strains with slt-II sequences, but did not react with any of the seven strains that carried slt-IIv, or slt-I and slt-IIv. The high SLT variability shown here has diagnostic implications and may well have consequences for the host response in infections associated with these pathogens.
authors.affiliation: Institut für Hygiene und Mikrobiologie, Universität Würzburg, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Rüssmann
authors.forename: H
authors.initials: H
authors.lastname: Schmidt
authors.forename: J
authors.initials: J
authors.lastname: Heesemann
authors.forename: A
authors.initials: A
authors.lastname: Caprioli
authors.forename: H
authors.initials: H
authors.lastname: Karch
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D015139:Blotting, Southern; D002648:Child; D017931:DNA Primers / Q000737:chemistry; D004768:Enterotoxins / Q000096:biosynthesis* / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D004927:Escherichia coli Infections / Q000150:complications / Q000382:microbiology*; D015964:Gene Expression Regulation, Bacterial; D005838:Genotype; D006463:Hemolytic-Uremic Syndrome / Q000209:etiology / Q000382:microbiology*; D006801:Humans; D008969:Molecular Sequence Data; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D022641:Shiga Toxin 2; D014709:Vero Cells
chemical_list: D001427:Bacterial Toxins; D017931:DNA Primers; D004768:Enterotoxins; D022641:Shiga Toxin 2