title: Development and evaluation of genotypic assays for the detection and characterization of enterotoxigenic Escherichia coli.
abstract: We developed and evaluated a method to genotypically identify enterotoxigenic Escherichia coli (ETEC) and to characterize these organisms with respect to 18 of 21 known colonization factors (CFs). The method, which is based on polynucleotide DNA-DNA colony hybridization, includes a pooled toxin probe assay to identify ETEC, and individual probe assays to detect the enterotoxins STp, STh, and LT, and the CFs CFA/I, CS1-CS8, CS12-CS15, CS17-CS19, CS21, and CS22. We evaluated the pooled toxin probe assay during a cohort study of childhood diarrhea, and the individual probe assays against 33 reference strains and 92 clinical ETEC isolates. There was close to a complete agreement between the pooled toxin probe assay and the individual toxin probe assays, and between the individual CF probe assays and the corresponding phenotypic assays.
authors.affiliation: Centre for International Health, University of Bergen, Norway. hans.steinsland@bio.uib.no
authors.forename: Hans
authors.initials: H
authors.lastname: Steinsland
authors.forename: Palle
authors.initials: P
authors.lastname: Valentiner-Branth
authors.forename: Harleen M S
authors.initials: HM
authors.lastname: Grewal
authors.forename: Wim
authors.initials: W
authors.lastname: Gaastra
authors.forename: Kåre
authors.initials: K
authors.lastname: Mølbak K
authors.forename: Halvor
authors.initials: H
authors.lastname: Sommerfelt
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D002648:Child; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis*; D003967:Diarrhea / Q000175:diagnosis*; D004768:Enterotoxins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D004927:Escherichia coli Infections / Q000175:diagnosis*; D005798:Genes, Bacterial; D005838:Genotype; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D015342:DNA Probes; D004269:DNA, Bacterial; D004768:Enterotoxins