title: Multicenter evaluation of a sequence-based protocol for subtyping Shiga toxins and standardizing Stx nomenclature.
abstract: When Shiga toxin-producing Escherichia coli (STEC) strains emerged as agents of human disease, two types of toxin were identified: Shiga toxin type 1 (Stx1) (almost identical to Shiga toxin produced by Shigella dysenteriae type 1) and the immunologically distinct type 2 (Stx2). Subsequently, numerous STEC strains have been characterized that express toxins with variations in amino acid sequence, some of which confer unique biological properties. These variants were grouped within the Stx1 or Stx2 type and often assigned names to indicate that they were not identical in sequence or phenotype to the main Stx1 or Stx2 type. A lack of specificity or consistency in toxin nomenclature has led to much confusion in the characterization of STEC strains. Because serious outcomes of infection have been attributed to certain Stx subtypes and less so with others, we sought to better define the toxin subtypes within the main Stx1 and Stx2 types. We compared the levels of relatedness of 285 valid sequence variants of Stx1 and Stx2 and identified common sequences characteristic of each of three Stx/Stx1 and seven Stx2 subtypes. A novel, simple PCR subtyping method was developed, independently tested on a battery of 48 prototypic STEC strains, and improved at six clinical and research centers to test the reproducibility, sensitivity, and specificity of the PCR. Using a consistent schema for nomenclature of the Stx toxins and stx genes by phylogenetic sequence-based relatedness of the holotoxin proteins, we developed a typing approach that should obviate the need to bioassay each newly described toxin and that predicts important biological characteristics.
authors: WHO Collaborating Centre for Reference and Research on Escherichia and Klebsiella, Unit of Foodborne Bacteria and Typing, Department of Microbiology and Infection Control, Statens Serum Institut, Copenhagen, Denmark. fsc@ssi.dk
authors: Flemming
authors: F
authors: Scheutz
authors: Louise D
authors: LD
authors: Teel
authors: Lothar
authors: L
authors: Beutin
authors: Denis
authors: D
authors: Piérard
authors: Glenn
authors: G
authors: Buvens
authors: Helge
authors: H
authors: Karch
authors: Alexander
authors: A
authors: Mellmann
authors: Alfredo
authors: A
authors: Caprioli
authors: Rosangela
authors: R
authors: Tozzoli
authors: Stefano
authors: S
authors: Morabito
authors: Nancy A
authors: NA
authors: Strockbine
authors: Angela R
authors: AR
authors: Melton-Celsa
authors: Maria
authors: M
authors: Sanchez
authors: Søren
authors: S
authors: Persson
authors: Alison D
authors: AD
authors: O'Brien
mesh_terms: D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D022603:Shiga Toxins / Q000145:classification* / Q000235:genetics*; D054323:Shiga-Toxigenic Escherichia coli / Q000235:genetics*; D009626:Terminology as Topic
chemical_list: D022603:Shiga Toxins