1887

Abstract

In this prospective multicentre study, an enzyme-linked fluorescent assay (VIDAS CDA2; bioMérieux), an enzyme-linked assay [Premier Toxins A and B (PTAB); Meridian] and an in-house real-time PCR amplifying the gene were compared with the cell cytotoxicity assay used as the ‘gold standard’ for diagnosis of -associated diarrhoea (CDAD). Faecal samples from patients with a request for diagnosis and samples from patients with diarrhoea hospitalized for at least 72 h were collected for 3 consecutive months from four university medical centres in The Netherlands. In total, 547 faecal samples were obtained from 450 patients. Of 540 samples available for all of the assays, 84 (15.6 %) showed a positive result in one or more assays. The cell cytotoxicity assay was positive in 31 samples (5.7 %) from 28 patients. A diagnosis of CDAD was not considered by the physician in 5 (23.8 %) of 21 patients with CDAD who were hospitalized for at least 72 h. Compared with the cell cytotoxicity assay, the sensitivity of VIDAS, PTAB and PCR was 83.9, 96.8 and 87.1 %, respectively. The specificity of VIDAS, PTAB and PCR was 97.1, 94.3 and 96.5 %, respectively. The positive and negative predictive values for VIDAS, PTAB and PCR were 63.4 and 99.0 %, 50.9 and 99.8 %, and 60.0 and 99.2 %, respectively. Of 61 samples that were positive in one, two or three of the assays, 56 were available for discordance analysis. Discordance analysis was performed by culture of toxinogenic strains. The concordance of VIDAS, PTAB and PCR with culture was 53.6 % (30/56), 55.4 % (31/56) and 71.4 % (40/56), respectively. It was concluded that real-time PCR had the highest concordance with toxinogenic culture and is therefore the preferred method for diagnosing CDAD in faecal samples. It was also concluded that diagnosis of patients with diarrhoea who have been hospitalized for more than 72 h should focus mainly on the detection of , irrespective of the physician's request.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.46680-0
2007-01-01
2019-11-17
Loading full text...

Full text loading...

/deliver/fulltext/jmm/56/1/36.html?itemId=/content/journal/jmm/10.1099/jmm.0.46680-0&mimeType=html&fmt=ahah

References

  1. Bowman, R. A., Bowman, J. M., Arrow, S. A. & Riley, T. V. ( 1992; ). Selective criteria for the microbiological examination of faecal specimens. J Clin Pathol 45, 838–839.[CrossRef]
    [Google Scholar]
  2. Brazier, J. S. ( 1998; ). The diagnosis of Clostridium difficile-associated disease. J Antimicrob Chemother 41, 29–40.
    [Google Scholar]
  3. Brown, E., Talbot, G. H., Axelrod, P., Provencher, M. & Hoegg, C. ( 1990; ). Risk factors for Clostridium difficile toxin-associated diarrhea. Infect Control Hosp Epidemiol 11, 283–290.[CrossRef]
    [Google Scholar]
  4. Clabots, C. R., Johnson, S., Olson, M. M., Peterson, L. R. & Gerding, D. N. ( 1992; ). Acquisition of Clostridium difficile by hospitalized patients: evidence for colonized new admissions as a source of infection. J Infect Dis 166, 561–567.[CrossRef]
    [Google Scholar]
  5. Delmée, M., van Broeck, J., Simon, A., Janssens, M. & Avesani, V. ( 2005; ). Laboratory diagnosis of Clostridium difficile-associated diarrhoea: a plea for culture. J Med Microbiol 54, 187–191.[CrossRef]
    [Google Scholar]
  6. Drudy, D., Harnedy, N., Fanning, S., O'Mahony, R., Baird, A. & Kyne, L. ( 2004; ). Endemic toxin variant Clostridium difficile in an Irish teaching hospital. In Abstracts of the First International C. difficile Symposium, 5–7 May 2004, Kranjska Gora, Slovenia, poster P2, p. 44. Edited by Maja Rupnik. Ljubljana.
  7. Fan, K., Morris, A. J. & Reller, L. B. ( 1993; ). Application of rejection criteria for stool cultures for bacterial enteric pathogens. J Clin Microbiol 31, 2233–2235.
    [Google Scholar]
  8. Freeman, J. & Wilcox, M. H. ( 2003; ). The effects of storage conditions on viability of Clostridium difficile vegetative cells and spores and toxin activity in human faeces. J Clin Pathol 56, 126–128.[CrossRef]
    [Google Scholar]
  9. García, A., García, T. & Pérez, J. L. ( 1997; ). Proline-aminopeptidase test for rapid screening of Clostridium difficile. J Clin Microbiol 35, 3007.
    [Google Scholar]
  10. Johnson, S. & Gerding, D. N. ( 1998; ). Clostridium difficile-associated diarrhea. Clin Infect Dis 26, 1027–1034.[CrossRef]
    [Google Scholar]
  11. Kato, H., Kato, N., Watanabe, K., Iwai, N., Nakamura, H., Yamamoto, T., Suzuki, K., Kim, S.-M., Chong, Y. & Wasito, E. B. ( 1998; ). Identification of toxin A-negative, toxin B-positive Clostridium difficile by PCR. J Clin Microbiol 36, 2178–2182.
    [Google Scholar]
  12. Kato, H., Kato, N., Katow, S., Maegawa, T., Nakamura, S. & Lyerly, D. M. ( 1999; ). Deletions in the repeating sequences of the toxin A gene of toxin A-negative, toxin B-positive Clostridium difficile strains. FEMS Microbiol Lett 175, 197–203.[CrossRef]
    [Google Scholar]
  13. Kelly, C. P. & LaMont, J. T. ( 1998; ). Clostridium difficile infection. Annu Rev Med 49, 375–390.[CrossRef]
    [Google Scholar]
  14. Lipson, S. M., Tortora, G., Tempone, A., Fedorko, D. P. & Spitzer, E. D. ( 2003; ). Rapid detection of Clostridium difficile in stool using the VIDASR C. difficile Toxin A II assay. Diagn Microbiol Infect Dis 45, 117–121.[CrossRef]
    [Google Scholar]
  15. O'Connor, D., Hynes, P., Cormican, M., Collins, E., Corbett-Feeney, G. & Cassidy, M. ( 2001; ). Evaluation of methods for detection of toxins in specimens of feces submitted for diagnosis of Clostridium difficile-associated diarrhea. J Clin Microbiol 39, 2846–2849.[CrossRef]
    [Google Scholar]
  16. Oldfield, E. C., III ( 2004; ). Clostridium difficile-associated diarrhea: risk factors, diagnostic methods, and treatment. Rev Gastroenterol Disord 4, 186–195.
    [Google Scholar]
  17. Renshaw, A. A., Stelling, J. M. & Doolittle, M. H. ( 1996; ). The lack of value of repeated Clostridium difficile cytotoxicity assays. Arch Pathol Lab Med 120, 49–52.
    [Google Scholar]
  18. Siegel, D. L., Edelstein, P. H. & Nachamkin, I. ( 1990; ). Inappropriate testing for diarrheal diseases in the hospital. JAMA 263, 979–982.[CrossRef]
    [Google Scholar]
  19. Turgeon, D. K., Novicki, T. J., Quick, J., Carlson, L., Miller, P., Ulness, B., Cent, A., Ashley, R., Larson, A. & other authors ( 2003; ). Six rapid tests for direct detection of Clostridium difficile and its toxins in fecal samples compared with the fibroblast cytotoxicity assay. J Clin Microbiol 41, 667–670.[CrossRef]
    [Google Scholar]
  20. van den Berg, R. J., Claas, E. C. J., Oyib, D. H., Klaassen, C. H. W., Dijkshoorn, L., Brazier, J. S. & Kuijper, E. J. ( 2004; ). Characterization of toxin A-negative, toxin B-positive Clostridium difficile isolates from outbreaks in different countries by amplified fragment length polymorphism and PCR ribotyping. J Clin Microbiol 42, 1035–1041.[CrossRef]
    [Google Scholar]
  21. van den Berg, R. J., Ameen, H. A. A., Furusawa, T., Claas, E. C. J., van der Vorm, E. R. & Kuijper, E. J. ( 2005a; ). Coexistence of multiple PCR-ribotype strains of Clostridium difficile in faecal samples limits epidemiological studies. J Med Microbiol 54, 173–179.[CrossRef]
    [Google Scholar]
  22. van den Berg, R. J., Legaria, M. C., de Breij, A., van der Vorm, E. R., Brazier, J. S. & Kuijper, E. J. ( 2005b; ). Introduction of TcdA-negative, TcdB-positive Clostridium difficile in a general hospital in Argentina. Clin Microbiol Infect 11 (Suppl. 2), 488.
    [Google Scholar]
  23. van den Berg, R. J., Kuijper, E. J., van Coppenraet, L. E. & Claas, E. C. J. ( 2006; ). Rapid diagnosis of toxinogenic Clostridium difficile in faecal samples with internally controlled real-time PCR. Clin Microbiol Infect 12, 184–186.[CrossRef]
    [Google Scholar]
  24. Verity, P., Wilcox, M. H., Fawley, W. & Parnell, P. ( 2001; ). Prospective evaluation of environmental contamination by Clostridium difficile in isolation side rooms. J Hosp Infect 49, 204–209.[CrossRef]
    [Google Scholar]
  25. Wilcox, M. H., Fawley, W. N. & Parnell, P. ( 2000; ). Value of lysozyme agar incorporation and alkaline thioglycollate exposure for the environmental recovery of Clostridium difficile. J Hosp Infect 44, 65–69.[CrossRef]
    [Google Scholar]
  26. Yannelli, B., Gurevich, I., Schoch, P. E. & Cunha, B. A. ( 1988; ). Yield of stool cultures, ova and parasite tests, and Clostridium difficile determinations in nosocomial diarrheas. Am J Infect Control 16, 246–249.[CrossRef]
    [Google Scholar]
  27. Zheng, L., Keller, S. F., Lyerly, D. M., Carman, R. J., Genheimer, C. W., Gleaves, C. A., Kohlhepp, S. J., Young, S., Perez, S. & Ye, K. ( 2004; ). Multicenter evaluation of a new screening test that detects Clostridium difficile in fecal specimens. J Clin Microbiol 42, 3837–3840.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.46680-0
Loading
/content/journal/jmm/10.1099/jmm.0.46680-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error