Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1986 Sep;24(3):384–387. doi: 10.1128/jcm.24.3.384-387.1986

Immunoblotting to demonstrate antigenic and immunogenic differences among nine standard strains of Clostridium difficile.

S R Heard, B Rasburn, R C Matthews, S Tabaqchali
PMCID: PMC268919  PMID: 2428826

Abstract

The epidemiology of Clostridium difficile-associated disease is being elucidated with the development of typing schemes for the organism. We recently described a new typing scheme based on the incorporation of [35S]methionine into bacterial proteins followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Nine standard strains were identified. We report here some observations on the antigenic differences among these nine strains when studied by immunoblotting. Type-specific rabbit antiserum was raised against each of the nine standard strains. Immunoblotting of the strains with these antisera demonstrated, in addition to the presence of shared, common proteins, a type-specific response with homologous antisera. When [35S]methionine-labeled C. difficile proteins were immunoblotted with homologous and heterologous antisera, both the immunoblots and the autoradiographs demonstrated the same strain-specific response. These strain-specific proteins, which have been so useful for epidemiological and typing purposes, were also immunogenic.

Full text

PDF
384

Images in this article

385Image
on p.385
385Image
on p.385
386Image
on p.386
386Image
on p.386
386Image
on p.386
386Image
on p.386

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Delmee M., Homel M., Wauters G. Serogrouping of Clostridium difficile strains by slide agglutination. J Clin Microbiol. 1985 Mar;21(3):323–327. doi: 10.1128/jcm.21.3.323-327.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Heard S. R., O'Farrell S., Holland D., Crook S., Barnett M. J., Tabaqchali S. The epidemiology of Clostridium difficile with use of a typing scheme: nosocomial acquisition and cross-infection among immunocompromised patients. J Infect Dis. 1986 Jan;153(1):159–162. doi: 10.1093/infdis/153.1.159. [DOI] [PubMed] [Google Scholar]
  3. Larson H. E., Price A. B., Honour P., Borriello S. P. Clostridium difficile and the aetiology of pseudomembranous colitis. Lancet. 1978 May 20;1(8073):1063–1066. doi: 10.1016/s0140-6736(78)90912-1. [DOI] [PubMed] [Google Scholar]
  4. Nakamura S., Serikawa T., Mikawa M., Nakashio S., Yamakawa K., Nishida S. Agglutination, toxigenicity and sorbitol fermentation of Clostridium difficile. Microbiol Immunol. 1981;25(9):863–870. doi: 10.1111/j.1348-0421.1981.tb00091.x. [DOI] [PubMed] [Google Scholar]
  5. Poxton I. R., Aronsson B., Möllby R., Nord C. E., Collee J. G. Immunochemical fingerprinting of Clostridium difficile strains isolated from an outbreak of antibiotic-associated colitis and diarrhoea. J Med Microbiol. 1984 Jun;17(3):317–324. doi: 10.1099/00222615-17-3-317. [DOI] [PubMed] [Google Scholar]
  6. Rampling A., Warren R. E., Bevan P. C., Hoggarth C. E., Swirsky D., Hayhoe F. G. Clostridium difficile in haematological malignancy. J Clin Pathol. 1985 Apr;38(4):445–451. doi: 10.1136/jcp.38.4.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Sell T. L., Schaberg D. R., Fekety F. R. Bacteriophage and bacteriocin typing scheme for Clostridium difficile. J Clin Microbiol. 1983 Jun;17(6):1148–1152. doi: 10.1128/jcm.17.6.1148-1152.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Tabaqchali S., Holland D., O'Farrell S., Silman R. Typing scheme for Clostridium difficile: its application in clinical and epidemiological studies. Lancet. 1984 Apr 28;1(8383):935–938. doi: 10.1016/s0140-6736(84)92392-4. [DOI] [PubMed] [Google Scholar]
  9. Tabaqchali S., O'Farrell S., Holland D., Silman R. Method for the typing of Clostridium difficile based on polyacrylamide gel electrophoresis of [35S]methionine-labeled proteins. J Clin Microbiol. 1986 Jan;23(1):197–198. doi: 10.1128/jcm.23.1.197-198.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Towbin H., Gordon J. Immunoblotting and dot immunobinding--current status and outlook. J Immunol Methods. 1984 Sep 4;72(2):313–340. doi: 10.1016/0022-1759(84)90001-2. [DOI] [PubMed] [Google Scholar]
  11. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Viscidi R., Laughon B. E., Yolken R., Bo-Linn P., Moench T., Ryder R. W., Bartlett J. G. Serum antibody response to toxins A and B of Clostridium difficile. J Infect Dis. 1983 Jul;148(1):93–100. doi: 10.1093/infdis/148.1.93. [DOI] [PubMed] [Google Scholar]
  13. Wüst J., Sullivan N. M., Hardegger U., Wilkins T. D. Investigation of an outbreak of antibiotic-associated colitis by various typing methods. J Clin Microbiol. 1982 Dec;16(6):1096–1101. doi: 10.1128/jcm.16.6.1096-1101.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES