1993
DOI: 10.1128/jcm.31.2.413-415.1993
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Typing of Clostridium difficile by western immunoblotting with 10 different antisera

Abstract: Western blotting (immunoblotting) with antisera against each of 10 reference serogroups was evaluated as a means of typing Clostridium diJfficile. A total of 164 clinical isolates of C. difficik were tested. Variations in band profiles in each serogroup were used to type isolates into subserogroups. This technique was useful for an epidemiological investigation. Outbreaks of Clostridium difficile-associated diarrhea in hospitalized patients are well documented (3, 5, 7, 8, 10). The availability of a simple, ra… Show more

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Cited by 20 publications
(9 citation statements)
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“…All strains were positive for toxin B by cell culture assay and negative for both toxin A by ELISA using monoclonal antibody and an ileal loop test in rabbits as reported previously [13]. All strains except for one (GAI 95606, serogroup X) were of serogroup F [14]. A total of 50 toxin A3, toxin B+ C. di¤cile strains isolated in various clinical settings were examined by PCR assay to con¢rm the deletion of the repeating regions of the toxin A gene.…”
Section: Bacterial Strainsmentioning
confidence: 68%
See 1 more Smart Citation
“…All strains were positive for toxin B by cell culture assay and negative for both toxin A by ELISA using monoclonal antibody and an ileal loop test in rabbits as reported previously [13]. All strains except for one (GAI 95606, serogroup X) were of serogroup F [14]. A total of 50 toxin A3, toxin B+ C. di¤cile strains isolated in various clinical settings were examined by PCR assay to con¢rm the deletion of the repeating regions of the toxin A gene.…”
Section: Bacterial Strainsmentioning
confidence: 68%
“…Precipitant of supernatant with ammonium sulfate was harvested by centrifugation, redissolved in 6 M urea in TBS (10 mM Tris-HCl (pH 7.5), 150 mM NaCl), and dialyzed against TBS. The immunoblotting was performed as described previously [14]. The concentrated culture supernatant (15 Wg of total protein) and puri¢ed native toxin A were separated by 5% SDS-PAGE, electrophoretically transferred to a nitrocellulose membrane (Schleicher and Schuell, Dassel, Germany), and probed with a 1:500 diluted goat polyclonal antiserum against toxin A.…”
Section: Page and Immunoblottingmentioning
confidence: 99%
“…The C. difficile strains were analyzed by Western immunoblotting (11) and pulsed-field gel electrophoresis (PFGE) typing (12). Immunoblot typing was performed with EDTA-extracted proteins of bacterial cells, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, as described previously (11), with antisera against reference strains of the 10 different serogroups established by a slide agglutination test (7). For PFGE analysis, the DNA in the inserts was digested with SacII (New England Biolabs, Beverly, Mass.…”
Section: Methodsmentioning
confidence: 99%
“…Efforts to understand the epidemiology of this important nosocomial pathogen have resulted in a variety of approaches for developing useful typing techniques. These include serotyping, plasmid profiles, polyacrylamide gel electrophoresis, immunoblotting, and restriction endonuclease analysis (4,6,8,9,(11)(12)(13)15). Most of these techniques are based on comparison of electrophoresis gel banding patterns of proteins or nucleic acids and require labor-intensive reagent preparation and/or extraction procedures.…”
mentioning
confidence: 99%
“…dijficile strains and sample preparation. The forty-one strains examined in this study were from a group of 50 strains isolated from patients during a hospital outbreak of colitis and diarrhea and were previously described and typed by Western immunoblotting (8).…”
mentioning
confidence: 99%