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. 1990 Oct;28(10):2210–2214. doi: 10.1128/jcm.28.10.2210-2214.1990

Characterization of flagella of Clostridium difficile and their role in serogrouping reactions.

M Delmée 1, V Avesani 1, N Delferriere 1, G Burtonboy 1
PMCID: PMC268149  PMID: 2229343

Abstract

Slide agglutination with rabbit antisera allows the differentiation of 10 serogroups of Clostridium difficile, namely, A, B, C, D, F, G, H, I, K, and X. Each serogroup displays a specific protein profile in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, except for A, which displays 12 different protein profiles (A1 to A12). In the present work, electron microscopy revealed the presence of uniformly distributed flagella in the reference strains of serogroups G and K and in all strains representative of the 12 subgroups within serogroup purified by differential centrifugation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of these preparations revealed one distinct band with an apparent molecular mass of approximately 39 kilodaltons. Antiserum was prepared by immunizing a rabbit with the serogroup A flagellin, which had been eluted from the gel. In immunoblotting, this antiserum cross-reacted with the flagellin of the other strains. When the cells were deflagellated by a short sonication, the cross-reactions observed by slide agglutination with A, G, and K antisera were suppressed. Similarly, shearing of flagella allowed specific slide agglutination of the 12 subgroups of serogroup A.

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Selected References

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

  1. Bacon A. E., Fekety R., Schaberg D. R., Faix R. G. Epidemiology of Clostridium difficile colonization in newborns: results using a bacteriophage and bacteriocin typing system. J Infect Dis. 1988 Aug;158(2):349–354. doi: 10.1093/infdis/158.2.349. [DOI] [PubMed] [Google Scholar]
  2. Clabots C. R., Peterson L. R., Gerding D. N. Characterization of a nosocomial Clostridium difficile outbreak by using plasmid profile typing and clindamycin susceptibility testing. J Infect Dis. 1988 Oct;158(4):731–736. doi: 10.1093/infdis/158.4.731. [DOI] [PubMed] [Google Scholar]
  3. Degl'Innocenti R., De Santis M., Berdondini I., Dei R. Outbreak of Clostridium difficile diarrhoea in an orthopaedic unit: evidence by phage-typing for cross-infection. J Hosp Infect. 1989 Apr;13(3):309–314. doi: 10.1016/0195-6701(89)90012-1. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Delmée M., Avesani V., Ernest I., Surleraux M. Detection of specific antigens for ten serogroups of Clostridium difficile. Mol Cell Probes. 1990 Feb;4(1):1–10. doi: 10.1016/0890-8508(90)90034-w. [DOI] [PubMed] [Google Scholar]
  6. Delmée M., Laroche Y., Avesani V., Cornelis G. Comparison of serogrouping and polyacrylamide gel electrophoresis for typing Clostridium difficile. J Clin Microbiol. 1986 Dec;24(6):991–994. doi: 10.1128/jcm.24.6.991-994.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Delmée M., Vandercam B., Avesani V., Michaux J. L. Epidemiology and prevention of Clostridium difficile infections in a leukemia unit. Eur J Clin Microbiol. 1987 Dec;6(6):623–627. doi: 10.1007/BF02013056. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Heard S. R., Rasburn B., Matthews R. C., Tabaqchali S. Immunoblotting to demonstrate antigenic and immunogenic differences among nine standard strains of Clostridium difficile. J Clin Microbiol. 1986 Sep;24(3):384–387. doi: 10.1128/jcm.24.3.384-387.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kelson J. S., Adler B., Chapman A. J., Faine S. Identification of leptospiral flagellar antigens by gel electrophoresis and immunoblotting. J Med Microbiol. 1988 May;26(1):47–53. doi: 10.1099/00222615-26-1-47. [DOI] [PubMed] [Google Scholar]
  11. Kodaka H., Armfield A. Y., Lombard G. L., Dowell V. R., Jr Practical procedure for demonstrating bacterial flagella. J Clin Microbiol. 1982 Nov;16(5):948–952. doi: 10.1128/jcm.16.5.948-952.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kuijper E. J., Oudbier J. H., Stuifbergen W. N., Jansz A., Zanen H. C. Application of whole-cell DNA restriction endonuclease profiles to the epidemiology of Clostridium difficile-induced diarrhea. J Clin Microbiol. 1987 Apr;25(4):751–753. doi: 10.1128/jcm.25.4.751-753.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. McFarland L. V., Mulligan M. E., Kwok R. Y., Stamm W. E. Nosocomial acquisition of Clostridium difficile infection. N Engl J Med. 1989 Jan 26;320(4):204–210. doi: 10.1056/NEJM198901263200402. [DOI] [PubMed] [Google Scholar]
  14. Mulligan M. E., Halebian S., Kwok R. Y., Cheng W. C., Finegold S. M., Anselmo C. R., Gerding D. N., Peterson L. R. Bacterial agglutination and polyacrylamide gel electrophoresis for typing Clostridium difficile. J Infect Dis. 1986 Feb;153(2):267–271. doi: 10.1093/infdis/153.2.267. [DOI] [PubMed] [Google Scholar]
  15. Mulligan M. E., Peterson L. R., Kwok R. Y., Clabots C. R., Gerding D. N. Immunoblots and plasmid fingerprints compared with serotyping and polyacrylamide gel electrophoresis for typing Clostridium difficile. J Clin Microbiol. 1988 Jan;26(1):41–46. doi: 10.1128/jcm.26.1.41-46.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pantosti A., Cerquetti M., Gianfrilli P. M. Electrophoretic characterization of Clostridium difficile strains isolated from antibiotic-associated colitis and other conditions. J Clin Microbiol. 1988 Mar;26(3):540–543. doi: 10.1128/jcm.26.3.540-543.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Peel M., Donachie W., Shaw A. Temperature-dependent expression of flagella of Listeria monocytogenes studied by electron microscopy, SDS-PAGE and western blotting. J Gen Microbiol. 1988 Aug;134(8):2171–2178. doi: 10.1099/00221287-134-8-2171. [DOI] [PubMed] [Google Scholar]
  18. Peerbooms P. G., Kuijt P., Maclaren D. M. Application of chromosomal restriction endonuclease digest analysis for use as typing method for Clostridium difficile. J Clin Pathol. 1987 Jul;40(7):771–776. doi: 10.1136/jcp.40.7.771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Steinberg J. P., Beckerdite M. E., Westenfelder G. O. Plasmid profiles of Clostridium difficile isolates from patients with antibiotic-associated colitis in two community hospitals. J Infect Dis. 1987 Dec;156(6):1036–1038. doi: 10.1093/infdis/156.6.1036. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Tanaka M., Hirayama N., Tamura Y. Production, characterization, and protective effect of monoclonal antibodies to Clostridium chauvoei flagella. Infect Immun. 1987 Aug;55(8):1779–1783. doi: 10.1128/iai.55.8.1779-1783.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Testore G. P., Pantosti A., Cerquetti M., Babudieri S., Panichi G., Gianfrilli P. M. Evidence for cross-infection in an outbreak of Clostridium difficile-associated diarrhoea in a surgical unit. J Med Microbiol. 1988 Jun;26(2):125–128. doi: 10.1099/00222615-26-2-125. [DOI] [PubMed] [Google Scholar]
  24. Toma S., Lesiak G., Magus M., Lo H. L., Delmée M. Serotyping of Clostridium difficile. J Clin Microbiol. 1988 Mar;26(3):426–428. doi: 10.1128/jcm.26.3.426-428.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Wren B. W., Tabaqchali S. Restriction endonuclease DNA analysis of Clostridium difficile. J Clin Microbiol. 1987 Dec;25(12):2402–2404. doi: 10.1128/jcm.25.12.2402-2404.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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