title: Detection by ELISA of low numbers of Shiga-like toxin-producing Escherichia coli in mixed cultures after growth in the presence of mitomycin C.
abstract: Techniques currently available to detect Shiga-like toxin (SLT)-producing Escherichia coli lack sensitivity or require specialised equipment and facilities, and in some cases detect only strains belonging to serotype O157. We have used an ELISA technique, capable of detecting both SLTI and SLTII with crude P1 glycoprotein from hydatid cysts, in combination with enhancement of toxin production by culture with mitomycin C. Supernates of Tryptone Soya Broth cultures containing mitomycin C 200 ng/ml were tested for SLTII. For SLTI, cell lysates pre-treated with polymyxin B were tested. In tests with E. coli O157:H7 in mixed culture with E. coli strain C600 alone, or with E. coli C600, Proteus mirabilis and Enterococcus faecalis, SLTI could be detected when the proportion of toxigenic organisms represented 1% of the mixture, and SLTII when the proportion was 0.025%. When faecal samples with added E. coli O157:H7 were examined in this system, SLTII-producing strains were detected when they comprised less than 0.1% of the coliform population. This technique is a sensitive and specific assay for detecting low numbers of SLT-producing organisms in mixed culture such as occurs in cases of haemolytic uraemic syndrome and haemorrhagic colitis.
authors.affiliation: Department of Microbiology, Hope Hospital, Salford, Manchester.
authors.forename: D
authors.initials: D
authors.lastname: Law
authors.forename: L A
authors.initials: LA
authors.lastname: Ganguli
authors.forename: A
authors.initials: A
authors.lastname: Donohue-Rolfe
authors.forename: D W
authors.initials: DW
authors.lastname: Acheson
mesh_terms: D001427:Bacterial Toxins / Q000032:analysis / Q000096:biosynthesis*; D013293:Enterococcus faecalis / Q000254:growth & development; D004768:Enterotoxins / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism; D004927:Escherichia coli Infections / Q000175:diagnosis; D005243:Feces / Q000382:microbiology; D006801:Humans; D016685:Mitomycin / Q000494:pharmacology*; D011237:Predictive Value of Tests; D011513:Proteus mirabilis / Q000254:growth & development; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2
chemical_list: D001427:Bacterial Toxins; D004768:Enterotoxins; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2; D016685:Mitomycin