1887

Abstract

A multilocus sequence analysis of ten virulence-associated genes was performed to study the genetic relationships between 29 isolates of various origins, hosts and clinical presentations, and selected from the main lineages previously defined by multilocus sequence typing (MLST) of housekeeping genes. Colonization-factor-encoding genes (, , , , , and ), toxin A and B genes ( and ), and the toxin A and B positive regulator gene () were investigated. Binary toxin genes ( and ) were also detected, and internal fragments were sequenced for positive isolates. Virulence-associated genes exhibited a moderate polymorphism, comparable to the polymorphism of housekeeping genes, whereas and genes appeared highly polymorphic. Isolates recovered from human pseudomembranous colitis cases did not define a specific lineage. The presence of binary toxin genes, detected in five of the 29 isolates (17 %), was also not linked to clinical presentation. Conversely, toxigenic A−B+ isolates defined a very homogeneous lineage, which is distantly related to other isolates. By clustering analysis, animal isolates were intermixed with human isolates. Multilocus sequence analysis of virulence-associated genes is consistent with a clonal population structure for and with the lack of host specificity. The data suggest a co-evolution of several of the virulence-associated genes studied (including toxins A and B and the binary toxin genes) with housekeeping genes, reflecting the genetic background of , whereas flagellin, and genes may undergo recombination events and/or environmental selective pressure.

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2005-10-01
2020-04-06
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References

  1. Alfa M. J., Kabani A., Lyerly D. 7 other authors 2000; Characterization of a toxin A-negative, toxin B-positive strain of Clostridium difficile responsible for a nosocomial outbreak of Clostridium difficile -associated diarrhea. J Clin Microbiol38:2706–2714
    [Google Scholar]
  2. Arroyo L. G., Kruth S. A., Willey B. M., Staempfli H. R., Low D. E., Weese J. S. 2005; PCR ribotyping of Clostridium difficile isolates originating from human and animal sources. J Med Microbiol54:163–166[CrossRef]
    [Google Scholar]
  3. Barbut F., Mario N., Meyohas M. C., Binet D., Frottier J., Petit J. C. 1994; Investigation of a nosocomial outbreak of Clostridium difficile -associated diarrhoea among AIDS patients by random amplified polymorphic DNA (RAPD) assay. J Hosp Infect26:181–189[CrossRef]
    [Google Scholar]
  4. Barbut F., Decre D., Lalande V. 9 other authors 2005; Clinical features of Clostridium difficile -associated diarrhoea due to binary toxin (actin-specific ADP-ribosyltransferase)-producing strains. J Med Microbiol54:181–185[CrossRef]
    [Google Scholar]
  5. Bartlett J. G. 1994; Clostridium difficile : history of its role as an enteric pathogen and the current state of knowledge about the organism. Clin Infect Dis18:S265–S272[CrossRef]
    [Google Scholar]
  6. Bartlett J. G., Moon N., Chang T. W., Taylor N., Onderdonk A. B. 1978; Role of Clostridium difficile in antibiotic-associated pseudomembranous colitis. Gastroenterology75:778–782
    [Google Scholar]
  7. Bidet P., Lalande V., Salauze B., Burghoffer B., Avesani V., Delmee M., Rossier A., Barbut F., Petit J. C. 2000; Comparison of PCR-ribotyping, arbitrarily primed PCR, and pulsed-field gel electrophoresis for typing Clostridium difficile . J Clin Microbiol38:2484–2487
    [Google Scholar]
  8. Bongaerts G. P., Lyerly D. M. 1994; Role of toxins A and B in the pathogenesis of Clostridium difficile disease. Microb Pathog17:1–12[CrossRef]
    [Google Scholar]
  9. Borriello S. P. 1998; Pathogenesis of Clostridium difficile infection. J Antimicrob Chemother41:13–19
    [Google Scholar]
  10. Borriello S. P., Ketley J. M., Mitchell T. J., Barclay F. E., Welch A. R., Price A. B., Stephen J. 1987; Clostridium difficile – a spectrum of virulence and analysis of putative virulence determinants in the hamster model of antibiotic-associated colitis. J Med Microbiol24:53–64[CrossRef]
    [Google Scholar]
  11. Borriello S. P., Wren B. W., Hyde S., Seddon S. V., Sibbons P., Krishna M. M., Tabaqchali S., Manek S., Price A. B. 1992; Molecular, immunological, and biological characterization of a toxin A-negative, toxin B-positive strain of Clostridium difficile . Infect Immun60:4192–4199
    [Google Scholar]
  12. Brazier J. S., Stubbs S. L., Duerden B. I. 1999; Prevalence of toxin A negative/B positive Clostridium difficile strains. J Hosp Infect42:248–249
    [Google Scholar]
  13. Calabi E., Ward S., Wren B., Paxton T., Panico M., Morris H., Dell A., Dougan G., Fairweather N. 2001; Molecular characterization of the surface layer proteins from Clostridium difficile . Mol Microbiol40:1187–1199[CrossRef]
    [Google Scholar]
  14. Calabi E., Calabi F., Phillips A. D., Fairweather N. F. 2002; Binding of Clostridium difficile surface layer proteins to gastrointestinal tissues. Infect Immun70:5770–5778[CrossRef]
    [Google Scholar]
  15. Cartmill T. D., Panigrahi H., Worsley M. A., McCann D. C., Nice C. N., Keith E. 1994; Management and control of a large outbreak of diarrhoea due to Clostridium difficile . J Hosp Infect27:1–15[CrossRef]
    [Google Scholar]
  16. Cerquetti M., Molinari A., Sebastianelli A., Diociaiuti M., Petruzzelli R., Capo C., Mastrantonio P. 2000; Characterization of surface layer proteins from different Clostridium difficile clinical isolates. Microb Pathog28:363–372[CrossRef]
    [Google Scholar]
  17. Delmee M., Homel M., Wauters G. 1985; Serogrouping of Clostridium difficile strains by slide agglutination. J Clin Microbiol21:323–327
    [Google Scholar]
  18. Dhalluin A., Pestel-Caron M., Mory F., Leluan G., Lemeland J. F., Pons J. L, Lemée L.. 2003; Genotypic differentiation of twelve Clostridium species by polymorphism analysis of the triosephosphate isomerase ( tpi ) gene. Syst Appl Microbiol26:90–96[CrossRef]
    [Google Scholar]
  19. Eveillard M., Fourel V., Barc M. C., Kerneis S., Coconnier M. H., Karjalainen T., Bourlioux P., Servin A. L. 1993; Identification and characterization of adhesive factors of Clostridium difficile involved in adhesion to human colonic enterocyte-like Caco-2 and mucus-secreting HT29 cells in culture. Mol Microbiol7:371–381[CrossRef]
    [Google Scholar]
  20. Hennequin C., Porcheray F., Waligora-Dupriet A., Collignon A., Barc M., Bourlioux P., Karjalainen T. 2001; GroEL (Hsp60) of Clostridium difficile is involved in cell adherence. Microbiology147:87–96
    [Google Scholar]
  21. Hennequin C., Janoir C., Barc M. C., Collignon A., Karjalainen T. 2003; Identification and characterization of a fibronectin-binding protein from Clostridium difficile . Microbiology149:2779–2787[CrossRef]
    [Google Scholar]
  22. Johnson S., Gerding D. N. 1998; Clostridium difficile -associated diarrhea. Clin Infect Dis26:1027–1034[CrossRef]
    [Google Scholar]
  23. Karjalainen T., Saumier N., Barc M. C., Delmee M., Collignon A. 2002; Clostridium difficile genotyping based on slpA variable region in S-layer gene sequence: an alternative to serotyping. J Clin Microbiol40:2452–2458[CrossRef]
    [Google Scholar]
  24. Kato H., Kato N., Watanabe K. 7 other authors 1998; Identification of toxin A-negative, toxin B-positive Clostridium difficile by PCR. J Clin Microbiol36:2178–2182
    [Google Scholar]
  25. Kelly C. P., Pothoulakis C., LaMont J. T. 1994; Clostridium difficile colitis. N Engl J Med330:257–262[CrossRef]
    [Google Scholar]
  26. Kumar S., Tamura K., Jakobsen I. B., Nei M. 2001; mega2: molecular evolutionary genetics analysis software. Bioinformatics17:1244–1245[CrossRef]
    [Google Scholar]
  27. Kyne L., Warny M., Qamar A., Kelly C. P. 2000; Asymptomatic carriage of Clostridium difficile and serum levels of IgG antibody against toxin A. N Engl J Med342:390–397[CrossRef]
    [Google Scholar]
  28. Kyne L., Warny M., Qamar A., Kelly C. P. 2001; Association between antibody response to toxin A and protection against recurrent Clostridium difficile diarrhoea. Lancet357:189–193[CrossRef]
    [Google Scholar]
  29. Larson H. E., Price A. B., Honour P., Borriello S. P. 1978; Clostridium difficile and the aetiology of pseudomembranous colitis. Lancet1:1063–1066
    [Google Scholar]
  30. Lemée L., Dhalluin A., Pestel-Caron M., Lemeland J. F., Pons J. L. 2004a; Multilocus sequence typing analysis of human and animal Clostridium difficile isolates of various toxigenic types. J Clin Microbiol42:2609–2617[CrossRef]
    [Google Scholar]
  31. Lemée L., Dhalluin A., Testelin S., Mattrat M. A., Maillard K., Lemeland J. F., Pons J. L. 2004b; Multiplex PCR targeting tpi (triose phosphate isomerase), tcdA (toxin A), and tcdB (toxin B) genes for toxigenic culture of Clostridium difficile . J Clin Microbiol42:5710–5714[CrossRef]
    [Google Scholar]
  32. Lyerly D. M., Neville L. M., Evans D. T. 7 other authors 1998; Multicenter evaluation of the Clostridium difficile TOX A/B TEST. J Clin Microbiol36:184–190
    [Google Scholar]
  33. Mani N., Dupuy B. 2001; Regulation of toxin synthesis in Clostridium difficile by an alternative RNA polymerase sigma factor. Proc Natl Acad Sci U S A98:5844–5849[CrossRef]
    [Google Scholar]
  34. McEllistrem M. C., Carman R. J., Gerding D. N., Genheimer C. W., Zheng L. 2005; A hospital outbreak of Clostridium difficile disease associated with isolates carrying binary toxin genes. Clin Infect Dis40:265–272[CrossRef]
    [Google Scholar]
  35. Moncrief J. S., Barroso L. A., Wilkins T. D. 1997; Positive regulation of Clostridium difficile toxins. Infect Immun65:1105–1108
    [Google Scholar]
  36. O'Neill G., Adams J. E., Bowman R. A., Riley T. V. 1993; A molecular characterization of Clostridium difficile isolates from humans, animals and their environments. Epidemiol Infect111:257–264[CrossRef]
    [Google Scholar]
  37. Pituch H., van den Braak N., van Leeuwen W., van Belkum A., Martirosian G., Obuch-Woszczatynski P., Luczak M., Meisel-Mikolajczyk F. 2001; Clonal dissemination of a toxin-A-negative/toxin-B-positive Clostridium difficile strain from patients with antibiotic-associated diarrhea in Poland. Clin Microbiol Infect7:442–446[CrossRef]
    [Google Scholar]
  38. Poilane I., Karjalainen T., Barc M. C., Bourlioux P., Collignon A. 1998; Protease activity of Clostridium difficile strains. Can J Microbiol44:157–161[CrossRef]
    [Google Scholar]
  39. Rupnik M., Avesani V., Janc M., von Eichel-Streiber C., Delmee M. 1998; A novel toxinotyping scheme and correlation of toxinotypes with serogroups of Clostridium difficile isolates. J Clin Microbiol36:2240–2247
    [Google Scholar]
  40. Rupnik M., Brazier J. S., Duerden B. I., Grabnar M., Stubbs S. L. 2001; Comparison of toxinotyping and PCR ribotyping of Clostridium difficile strains and description of novel toxinotypes. Microbiology147:439–447
    [Google Scholar]
  41. Rupnik M., Kato N., Grabnar M., Kato H. 2003; New types of toxin A-negative, toxin B-positive strains among Clostridium difficile isolates from Asia. J Clin Microbiol41:1118–1125[CrossRef]
    [Google Scholar]
  42. Savariau-Lacomme M. P., Lebarbier C., Karjalainen T., Collignon A., Janoir C. 2003; Transcription and analysis of polymorphism in a cluster of genes encoding surface-associated proteins of Clostridium difficile . J Bacteriol185:4461–4470[CrossRef]
    [Google Scholar]
  43. Seddon S. V., Hemingway I., Borriello S. P. 1990; Hydrolytic enzyme production by Clostridium difficile and its relationship to toxin production and virulence in the hamster model. J Med Microbiol31:169–174[CrossRef]
    [Google Scholar]
  44. Smith J. M., Smith N. H., O'Rourke M., Spratt B. G. 1993; How clonal are bacteria?. Proc Natl Acad Sci U S A90:4384–4388[CrossRef]
    [Google Scholar]
  45. Spigaglia P., Mastrantonio P. 2002; Molecular analysis of the pathogenicity locus and polymorphism in the putative negative regulator of toxin production (TcdC) among Clostridium difficile clinical isolates. J Clin Microbiol40:3470–3475[CrossRef]
    [Google Scholar]
  46. Stubbs S. L., Brazier J. S., O'Neill G. L., Duerden B. I. 1999; PCR targeted to the 16S-23S rRNA gene intergenic spacer region of Clostridium difficile and construction of a library consisting of 116 different PCR ribotypes. J Clin Microbiol37:461–463
    [Google Scholar]
  47. Stubbs S., Rupnik M., Gibert M., Brazier J., Duerden B., Popoff M. 2000; Production of actin-specific ADP-ribosyltransferase (binary toxin) by strains of Clostridium difficile . FEMS Microbiol Lett186:307–312[CrossRef]
    [Google Scholar]
  48. Tasteyre A., Karjalainen T., Avesani V., Delmee M., Collignon A., Bourlioux P., Barc M. C. 2000; Phenotypic and genotypic diversity of the flagellin gene ( fliC ) among Clostridium difficile isolates from different serogroups. J Clin Microbiol38:3179–3186
    [Google Scholar]
  49. Tasteyre A., Barc M. C., Collignon A., Boureau H., Karjalainen T. 2001; Role of FliC and FliD flagellar proteins of Clostridium difficile in adherence and gut colonization. Infect Immun69:7937–7940[CrossRef]
    [Google Scholar]
  50. Toyokawa M., Ueda A., Tsukamoto H., Nishi I., Horikawa M., Sunada A., Asari S. 2003; Pseudomembranous colitis caused by toxin A-negative/toxin B-positive variant strain of Clostridium difficile . J Infect Chemother9:351–354[CrossRef]
    [Google Scholar]
  51. Waligora A. J., Hennequin C., Mullany P., Bourlioux P., Collignon A., Karjalainen T. 2001; Characterization of a cell surface protein of Clostridium difficile with adhesive properties. Infect Immun69:2144–2153[CrossRef]
    [Google Scholar]
  52. Zhang W., Jayarao B. M., Knabel S. J. 2004; Multi-virulence-locus sequence typing of Listeria monocytogenes . Appl Environ Microbiol70:913–920[CrossRef]
    [Google Scholar]
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