1887

Abstract

Previous reports have documented that a surface layer protein (SlpA) varies among isolates. The typing system by sequencing the variable region of the gene was applied to typing strains belonging to one PCR ribotype, type smz, which has been identified as frequently causing outbreaks in Japan. The PCR ribotype smz strains recovered from patients at different hospitals in Japan were examined. Among 10 type smz strains tested, three subtypes, smz-1, -2 and -3, were identified that differed from each other by one nucleotide. sequence typing was also applied to direct typing on DNA extracted from stool specimens. Of 22 stool specimens examined, 17 were PCR positive for ; eight were typed as sequence type smz-1 and nine as type smz-2. was cultured from 12 of these 17 stool specimens, and the sequence results of the recovered isolates were compared with those from the DNA extracted from the stool specimens. In all 12 of these stool specimens, the sequence results of DNA from recovered isolates completely agreed with those of DNA extracted directly from stool specimens. The remaining five stool specimens were culture-negative for . Sequence typing has the advantage of enabling easy comparison of typing results among multiple laboratories via the Internet without exchanging reference strains as is required in typing systems which depend on banding-pattern analyses. sequence typing appears to be a reproducible and reliable typing system for as well as being useful for the typing of when stool specimens contain only small numbers of or are inappropriate for culturing.

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2005-02-01
2020-01-24
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References

  1. Brazier, J. S. ( 1998;). The epidemiology and typing of Clostridium difficile. J Antimicrob Chemother 41 Suppl C, 47–57.
    [Google Scholar]
  2. Brazier, J. S., Mulligan, M. E., Delmee, M., Tabaqchali, S. & the International Clostridium difficile Study Group ( 1997;). Preliminary findings of the international typing study on Clostridium difficile. Clin Infect Dis 25, S199–S201.[CrossRef]
    [Google Scholar]
  3. 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 Microbiol 40, 1187–1199.[CrossRef]
    [Google Scholar]
  4. Corkill, J. E., Graham, R., Hart, C. A. & Stubbs, S. ( 2000;). Pulsed-field gel electrophoresis of degradation-sensitive DNAs from Clostridium difficile PCR ribotype 1 strains. J Clin Microbiol 38, 2791–2792.
    [Google Scholar]
  5. Johnson, S., Samore, M. H., Farrow, K. A. & 9 other authors ( 1999;). Epidemics of diarrhea caused by a clindamycin-resistant strain of Clostridium difficile in four hospitals. N Engl J Med 341, 1645–1651.[CrossRef]
    [Google Scholar]
  6. 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 Microbiol 40, 2452–2458.[CrossRef]
    [Google Scholar]
  7. Kato, H., Cavallaro, J. J., Kato, N., Bartley, S. L., Killgore, G. E., Watanabe, K. & Ueno, K. ( 1993;). Typing of Clostridium difficile by western immunoblotting with 10 different antisera. J Clin Microbiol 31, 413–415.
    [Google Scholar]
  8. Kato, N., Ou, C. Y., Kato, H., Bartley, S. L., Luo, C. C., Killgore, G. E. & Ueno, K. ( 1993;). Detection of toxigenic Clostridium difficile in stool specimens by the polymerase chain reaction. J Infect Dis 167, 455–458.[CrossRef]
    [Google Scholar]
  9. Kato, H., Kato, N., Watanabe, K. & 7 other authors ( 1998;). Identification of toxin A-negative, toxin B-positive Clostridium difficile by PCR. J Clin Microbiol 36, 2178–2182.
    [Google Scholar]
  10. 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]
  11. Kato, H., Kato, N., Watanabe, K. & 7 other authors ( 2001;). Analysis of Clostridium difficile isolates from nosocomial outbreaks at three hospitals in diverse areas of Japan. J Clin Microbiol 39, 1391–1395.[CrossRef]
    [Google Scholar]
  12. Killgore, G. E. & Kato, H. ( 1994;). Use of arbitrary primer PCR to type Clostridium difficile and comparison of results with those by immunoblot typing. J Clin Microbiol 32, 1591–1593.
    [Google Scholar]
  13. Lemee, L., Dhalluin, A., Pestel-Caron, M., Lemeland, J. F. & Pons, J. L. ( 2004;). Multilocus sequence typing analysis of human and animal Clostridium difficile isolates of various toxigenic types. J Clin Microbiol 42, 2609–2617.[CrossRef]
    [Google Scholar]
  14. McCoubrey, J. & Poxton, I. R. ( 2001;). Variation in the surface layer proteins of Clostridium difficile. FEMS Immunol Med Microbiol 31, 131–135.[CrossRef]
    [Google Scholar]
  15. Samore, M., Killgore, G., Johnson, S. & 8 other authors ( 1997;). Multicenter typing comparison of sporadic and outbreak Clostridium difficile isolates from geographically diverse hospitals. J Infect Dis 176, 1233–1238.[CrossRef]
    [Google Scholar]
  16. 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 Microbiol 37, 461–463.
    [Google Scholar]
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