title: Identification of Shiga-like toxin type II producing Escherichia coli using the polymerase chain reaction and a digoxigenin labelled DNA probe.
abstract: Epidemiological studies have demonstrated that enterohaemorrhagic strains of Escherichia coli which cause the haemolytic uremic syndrome in humans and the oedema disease in pigs more frequently produce Shiga-like toxin type II (SLT-II) than any other member of the Shiga-like toxin family. A technique has been developed for the identification of SLT-II producing E. coli using the polymerase chain reaction (PCR) and a digoxigenin (DIG)-labelled DNA probe to facilitate the early detection and epidemiological analysis of these pathogens. Whole cell DNA liberated from isolated colonies during the denaturation step of PCR was amplified using a primer pair which is homologous to the slt-II gene sequences. The amplification products were transferred directly to a nitrocellulose membrane or following agarose gel electrophoresis and DNA denaturation. A chemically labelled DNA probe, prepared using PCR with the incorporation of DIG, was used to identify the PCR products of strains which produced SLT-II or a variant of SLT-II.
authors.affiliation: Wayne State University School of Medicine, Department of Immunology and Microbiology, Detroit, MI 48201.
authors.forename: M P
authors.initials: MP
authors.lastname: Jackson
mesh_terms: D001427:Bacterial Toxins / Q000032:analysis* / Q000235:genetics; D015139:Blotting, Southern; D015342:DNA Probes; D004076:Digoxigenin; D004768:Enterotoxins / Q000032:analysis* / Q000235:genetics; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000175:diagnosis / Q000382:microbiology; D005798:Genes, Bacterial; D006801:Humans; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D022641:Shiga Toxin 2
chemical_list: D001427:Bacterial Toxins; D015342:DNA Probes; D004768:Enterotoxins; D022641:Shiga Toxin 2; D004076:Digoxigenin