Comparison of toxinotyping and PCR ribotyping of strains and description of novel toxinotypes Free

Abstract

Toxinotyping and PCR ribotyping are two methods that have been used to type isolates. Toxinotyping is based on PCR-RFLP analysis of a 19 kb region encompassing the pathogenicity locus. PCR ribotyping is based on comparison of patterns of PCR products of the 16S–23S rRNA intergenic spacer region. Representative strains (101) from a PCR ribotype library and 22 strains from previously described toxinotypes were analysed to compare ribotyping with toxinotyping. Within this panel of strains all 11 toxinotypes (0–X) described previously and an additional 5 novel toxinotypes (XI–XV) were observed. PCR ribotyping and toxinotyping correlated well and usually all strains within a given ribotype had similar changes in toxin genes. The new toxinotype XI comprises strains that did not express toxins TcdA or TcdB at detectable levels, but contained part of the gene. Strains of toxinotype XII exhibit changes only in the 5′ end of the gene. Toxinotype XIV is composed of strains that have a large insertion at the beginning of the gene. A total of 25 of the 89 tested PCR ribotypes of contained variant strains. It was estimated that they represent 77% of the total number of strains in the Anaerobe Reference Unit collection.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-2-439
2001-02-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/2/1470439a.html?itemId=/content/journal/micro/10.1099/00221287-147-2-439&mimeType=html&fmt=ahah

References

  1. Barbut F. N., Mario J., Frottier J., Petit J. C. 1993; Use of arbitrary primer PCR for investigating an outbreak of Clostridium difficile-associated diarrhoea in AIDS patients. Eur J Clin Microbiol Infect Dis 12:724–795
    [Google Scholar]
  2. 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 Immun 60:4192–4199
    [Google Scholar]
  3. Braun V., Hundsberger T., Leukel P., Sauerborn M., von Eichel-Streiber C. 1996; Definition of the single integration site of the pathogenicity locus in Clostridium difficile. Gene 181:29–38 [CrossRef]
    [Google Scholar]
  4. Braun V., Mehlig M., Moos M., Rupnik M., Kalt B., Mahony D. E., von Eichel-Streiber C. 2000; A chimeric ribozyme in Clostridium difficile combines features of group I introns and insertion elements. Mol Microbiol 36:1447–1459
    [Google Scholar]
  5. Brazier J. S. 1993; Role of the laboratory in investigations of Clostridium difficile diarrhea. Clin Infect Dis 16:(Suppl. 4)S228–S233
    [Google Scholar]
  6. Brazier J. S., Mulligan M. E., Delm é e M., Tabaqchali S. 1997; Preliminary findings of the international typing study on Clostridium difficile. International Clostridium Difficile Study Group. Clin Infect Dis 25:Suppl 2S199–S201 [CrossRef]
    [Google Scholar]
  7. Burdon D. W., Brown J. D., Youngs D. J., Arabi Y., Shinagawa N., Alexander-Williams J., Keinghley M. R. B. 1979; Antibiotic susceptibility of Clostridium difficile. J Antimicrob Chemother 5:307–310 [CrossRef]
    [Google Scholar]
  8. Cohen S. H., Tang Y. J., Hansen B., Silva J. Jr 1998; Isolation of a toxin B-deficient mutant strain of Clostridium difficile in a case of recurrent C. difficile-associated diarrhea. Clin Infect Dis 26:410–412 [CrossRef]
    [Google Scholar]
  9. Collier M. C., Stock F., DeGirolami P. C., Samore M. H., Cartwright C. P. 1996; Comparison of PCR-based approaches to molecular epidemiologic analysis of Clostridium difficile. J Clin Microbiol 34:1153–1157
    [Google Scholar]
  10. Delmée M., Homel M., Wauters G. 1985; Serogrouping of Clostridium difficile strains by slide agglutination. J Clin Microbiol 21:323–327
    [Google Scholar]
  11. Delmée M., Laroche Y., Avesani V., Cornelis G. 1986; Comparison of serogrouping and polyacrylamide gel electrophoresis for typing of Clostridium difficile. J Clin Microbiol 24:991–994
    [Google Scholar]
  12. Depitre C., Delmée M., Avesani V., L’Haridon R., Roels A., Popoff M., Corthier G. 1993; Serogroup F strains of Clostridium difficile produce toxin B but not toxin A. J Med Microbiol 38:434–441 [CrossRef]
    [Google Scholar]
  13. van Dijck P., Avesani V., Delmée M. 1996; Genotyping of outbreak-related and sporadic isolates of Clostridium difficile belonging to serogroup C. J Clin Microbiol 34:3049–3055
    [Google Scholar]
  14. von Eichel-Streiber C., Sauerborn M. 1990; Clostridium difficile toxin A carries a C-terminal repetitive structure homologous to the carbohydrate binding region of streptococcal glycosyltransferases. Gene 96:107–113 [CrossRef]
    [Google Scholar]
  15. von Eichel-Streiber C., Boquet P., Sauerborn M., Thelestam M. 1996; Large clostridial cytotoxins – a family of glycosyltransferases modifying small GTP-binding proteins. Trends Microbiol 4:375–382 [CrossRef]
    [Google Scholar]
  16. von Eichel-Streiber C., Zec-Pirnat I., Grabnar M., Rupnik M. 1999; A nonsense mutation abrogates production of functional enterotoxin A in Clostridium difficile toxinotype VIII strains of serogroups F and X. FEMS Microbiol Lett 178:163–168 [CrossRef]
    [Google Scholar]
  17. Hammond G. A., Johnson J. L. 1995; The toxinogenic element of Clostridium difficile strain VPI 10463. Microb Pathog 19:203–213 [CrossRef]
    [Google Scholar]
  18. Hutson R. A., Thompson D. E., Collins M. D. 1993; Genetic interrelationships of saccharolytic Clostridium botulinum types B, E and F and related clostridia as revealed by small-subunit rRNA gene sequences. FEMS Microbiol Lett 108:103–110 [CrossRef]
    [Google Scholar]
  19. Knoop F. C., Owens M., Crocker I. C. 1993; Clostridium difficile: clinical disease and diagnosis. Clin Microbiol Rev 6:251–265
    [Google Scholar]
  20. Lyerly D. M., Barroso L. A., Wilkins T. D., Depitre C., Corthier G. 1992; Characterization of a Toxin A-negative, Toxin B-positive strain of Clostridium difficile. Infect Immun 60:4633–4639
    [Google Scholar]
  21. Mathis J. N., Pilkinton L., McMillin D. E. 1999; Detection and transcription of toxin DNA in a nontoxigenic strain of Clostridium difficile. Curr Microbiol 38:324–328 [CrossRef]
    [Google Scholar]
  22. Mulligan M. E., Peterson L. R., Kwok R. Y. Y., Clabots C. R., Gerding D. N. 1988; Immunoblots and plasmid fingerprints compared with serotyping and polyacrylamide gel electrophoresis for typing Clostridium difficile. J Clin Microbiol 26:41–46
    [Google Scholar]
  23. O’Neill G. L., Beaman M. H., Riley T. V. 1991; Relapse versus reinfection with Clostridium difficile. Epidemiol Infect 107:627–635 [CrossRef]
    [Google Scholar]
  24. O’Neill G. L., Ogunsola F. T., Brazier J. S., Duerden B. I. 1996; Modification of a PCR rIbotyping method for application as a routine typing scheme for Clostridium difficile. Anaerobe 2:205–209 [CrossRef]
    [Google Scholar]
  25. Riley T. V. 1994; The epidemiology of Clostridium difficile – associated diarrhoea. Rev Med Microbiol 5:117–122 [CrossRef]
    [Google Scholar]
  26. Riley T. V. 1998; Clostridium difficile: a pathogen of the nineties. Eur J Clin Microbiol Infect Dis 17:137–141
    [Google Scholar]
  27. Rupnik M., Braun V., Soehn F., Janc M., Hofstetter M., Laufenberg-Feldman R., von Eichel-Streiber C. 1997; Characterization of polymorphisms in the toxin A and B genes of Clostridium difficile. FEMS Microbiol Lett 148:197–202 [CrossRef]
    [Google Scholar]
  28. Rupnik M., Avesani V., Janc M., von Eichel-Streiber C., Delmée M. 1998; A novel toxinotyping scheme and correlation of toxinotypes with serogroups of Clostridium difficile isolates. J Clin Microbiol 36:2240–2247
    [Google Scholar]
  29. Sell T. L., Schaberg D. R., Fekety F. R. 1983; Bacteriophage and bacteriocin typing scheme for Clostridium difficile. J Clin Microbiol 17:1148–1152
    [Google Scholar]
  30. Soehn F., Wagenknecht-Wiesner A., Leukel P., Kohl M., Weidman M., von Eichel-Streiber C., Braun V. 1998; Genetic rearrangements in the pathogenicity locus of Clostridium difficile strain 8864 – implications for transcription, expression and enzymatic activity of toxins A and B. Mol Gen Genet 258:222–232 [CrossRef]
    [Google Scholar]
  31. Stubbs S. L. J., 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]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-2-439
Loading
/content/journal/micro/10.1099/00221287-147-2-439
Loading

Data & Media loading...

Most cited Most Cited RSS feed