This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Bidet, P.
Right arrow Articles by Petit, J.-C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Bidet, P.
Right arrow Articles by Petit, J.-C.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, July 2000, p. 2484-2487, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Comparison of PCR-Ribotyping, Arbitrarily Primed PCR, and Pulsed-Field Gel Electrophoresis for Typing Clostridium difficile

Philippe Bidet,1,* Valérie Lalande,1 Béatrice Salauze,2 Béatrice Burghoffer,1 Véronique Avesani,3 Michel Delmée,3 Anne Rossier,2 Frédéric Barbut,1 and Jean-Claude Petit1

Laboratoire de Bactériologie, Hôpital Saint-Antoine,1 Laboratoire de Bactériologie, Hôpital Rothschild,2 Centre Hospitalo-Universitaire Saint-Antoine, Université Paris 6, Assistance Publique-Hôpitaux de Paris, Paris, France, and Laboratoire de Bactériologie, Université Catholique de Louvain, Bruxelles, Belgium3

Received 19 October 1999/Returned for modification 6 February 2000/Accepted 25 March 2000

Clostridium difficile is now recognized as the major agent responsible for nosocomial diarrhea in adults. Among the genotyping methods available, arbitrarily primed PCR (AP-PCR), PCR-ribotyping, and pulsed-field gel electrophoresis (PFGE) have been widely used for investigating outbreaks of C. difficile infections. However, the comparative typing ability, reproducibility, discriminatory power, and efficiency of these methods have not been fully investigated. We compared the results of three methods---AP-PCR with three different primers (AP3, AP4, and AP5), PCR-ribotyping, and PFGE (with SmaI endonuclease)---to differentiate 99 strains of C. difficile that had been previously serogrouped. Typing abilities were 100% for PCR-ribotyping and AP-PCR with AP3 and 90% for PFGE, due to early DNA degradation in strains from serogroup G. Reproducibilities were 100% for PCR-ribotyping and PFGE but only 88% for AP-PCR with AP3, 67% for AP-PCR with AP4, and 33% for AP-PCR with AP5. Discriminatory power for unrelated strains was >0.95 for all the methods but was lower for PCR-ribotyping among serogroups D and C. PCR-based methods were easier and quicker to perform, but their fingerprints were more difficult to interpret than those of PFGE. We conclude that PCR-ribotyping offers the best combination of advantages as an initial typing tool for C. difficile.

* Corresponding author. Present address: Laboratoire de Microbiologie, Hôpital Armand-Trousseau, 26 Ave. du Dr. Arnold Netter, 75012 Paris, France. Phone: 33 (1) 44 73 65 81. Fax: 33 (1) 44 73 62 88. E-mail: philippe.bidet{at}trs.ap-hop-paris.fr.

Journal of Clinical Microbiology, July 2000, p. 2484-2487, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Indra, A., Huhulescu, S., Schneeweis, M., Hasenberger, P., Kernbichler, S., Fiedler, A., Wewalka, G., Allerberger, F., Kuijper, E. J. (2008). Characterization of Clostridium difficile isolates using capillary gel electrophoresis-based PCR ribotyping. J Med Microbiol 57: 1377-1382 [Abstract] [Full Text]  
  • Martin, H., Willey, B., Low, D. E., Staempfli, H. R., McGeer, A., Boerlin, P., Mulvey, M., Weese, J. S. (2008). Characterization of Clostridium difficile Strains Isolated from Patients in Ontario, Canada, from 2004 to 2006. J. Clin. Microbiol. 46: 2999-3004 [Abstract] [Full Text]  
  • Pelaez, T., Cercenado, E., Alcala, L., Marin, M., Martin-Lopez, A., Martinez-Alarcon, J., Catalan, P., Sanchez-Somolinos, M., Bouza, E. (2008). Metronidazole Resistance in Clostridium difficile Is Heterogeneous. J. Clin. Microbiol. 46: 3028-3032 [Abstract] [Full Text]  
  • Indra, A., Schmid, D., Huhulescu, S., Hell, M., Gattringer, R., Hasenberger, P., Fiedler, A., Wewalka, G., Allerberger, F. (2008). Characterization of clinical Clostridium difficile isolates by PCR ribotyping and detection of toxin genes in Austria, 2006-2007. J Med Microbiol 57: 702-708 [Abstract] [Full Text]  
  • Bouchet, V., Huot, H., Goldstein, R. (2008). Molecular Genetic Basis of Ribotyping. Clin. Microbiol. Rev. 21: 262-273 [Abstract] [Full Text]  
  • Killgore, G., Thompson, A., Johnson, S., Brazier, J., Kuijper, E., Pepin, J., Frost, E. H., Savelkoul, P., Nicholson, B., van den Berg, R. J., Kato, H., Sambol, S. P., Zukowski, W., Woods, C., Limbago, B., Gerding, D. N., McDonald, L. C. (2008). Comparison of Seven Techniques for Typing International Epidemic Strains of Clostridium difficile: Restriction Endonuclease Analysis, Pulsed-Field Gel Electrophoresis, PCR-Ribotyping, Multilocus Sequence Typing, Multilocus Variable-Number Tandem-Repeat Analysis, Amplified Fragment Length Polymorphism, and Surface Layer Protein A Gene Sequence Typing. J. Clin. Microbiol. 46: 431-437 [Abstract] [Full Text]  
  • Mutlu, E., Wroe, A. J., Sanchez-Hurtado, K., Brazier, J. S., Poxton, I. R. (2007). Molecular characterization and antimicrobial susceptibility patterns of Clostridium difficile strains isolated from hospitals in south-east Scotland. J Med Microbiol 56: 921-929 [Abstract] [Full Text]  
  • Keel, K., Brazier, J. S., Post, K. W., Weese, S., Songer, J. G. (2007). Prevalence of PCR Ribotypes among Clostridium difficile Isolates from Pigs, Calves, and Other Species. J. Clin. Microbiol. 45: 1963-1964 [Abstract] [Full Text]  
  • Marsh, J. W., O'Leary, M. M., Shutt, K. A., Pasculle, A. W., Johnson, S., Gerding, D. N., Muto, C. A., Harrison, L. H. (2006). Multilocus Variable-Number Tandem-Repeat Analysis for Investigation of Clostridium difficile Transmission in Hospitals.. J. Clin. Microbiol. 44: 2558-2566 [Abstract] [Full Text]  
  • MacCannell, D. R., Louie, T. J., Gregson, D. B., Laverdiere, M., Labbe, A.-C., Laing, F., Henwick, S. (2006). Molecular Analysis of Clostridium difficile PCR Ribotype 027 Isolates from Eastern and Western Canada.. J. Clin. Microbiol. 44: 2147-2152 [Abstract] [Full Text]  
  • Leclair, D., Pagotto, F., Farber, J. M., Cadieux, B., Austin, J. W. (2006). Comparison of DNA fingerprinting methods for use in investigation of type e botulism outbreaks in the canadian arctic.. J. Clin. Microbiol. 44: 1635-1644 [Abstract] [Full Text]  
  • Akerlund, T., Svenungsson, B., Lagergren, A., Burman, L. G. (2006). Correlation of Disease Severity with Fecal Toxin Levels in Patients with Clostridium difficile-Associated Diarrhea and Distribution of PCR Ribotypes and Toxin Yields In Vitro of Corresponding Isolates. J. Clin. Microbiol. 44: 353-358 [Abstract] [Full Text]  
  • Pechine, S., Janoir, C., Collignon, A. (2005). Variability of Clostridium difficile Surface Proteins and Specific Serum Antibody Response in Patients with Clostridium difficile-Associated Disease. J. Clin. Microbiol. 43: 5018-5025 [Abstract] [Full Text]  
  • Lemee, L., Bourgeois, I., Ruffin, E., Collignon, A., Lemeland, J.-F., Pons, J.-L. (2005). Multilocus sequence analysis and comparative evolution of virulence-associated genes and housekeeping genes of Clostridium difficile. Microbiology 151: 3171-3180 [Abstract] [Full Text]  
  • Fawley, W. N., Parnell, P., Verity, P., Freeman, J., Wilcox, M. H. (2005). Molecular Epidemiology of Endemic Clostridium difficile Infection and the Significance of Subtypes of the United Kingdom Epidemic Strain (PCR Ribotype 1). J. Clin. Microbiol. 43: 2685-2696 [Abstract] [Full Text]  
  • Ohara-Nemoto, Y., Kishi, K., Satho, M., Tajika, S., Sasaki, M., Namioka, A., Kimura, S. (2005). Infective Endocarditis Caused by Granulicatella elegans Originating in the Oral Cavity. J. Clin. Microbiol. 43: 1405-1407 [Abstract] [Full Text]  
  • Alonso, R, Martin, A, Pelaez, T, Marin, M, Rodriguez-Creixems, M, Bouza, E (2005). An improved protocol for pulsed-field gel electrophoresis typing of Clostridium difficile. J Med Microbiol 54: 155-157 [Abstract] [Full Text]  
  • Alonso, R, Martin, A, Pelaez, T, Marin, M, Rodriguez-Creixems, M, Bouza, E (2005). Toxigenic status of Clostridium difficile in a large Spanish teaching hospital. J Med Microbiol 54: 159-162 [Abstract] [Full Text]  
  • Cartelle, M., del Mar Tomas, M., Pertega, S., Beceiro, A., Dominguez, M. A., Velasco, D., Molina, F., Villanueva, R., Bou, G. (2004). Risk Factors for Colonization and Infection in a Hospital Outbreak Caused by a Strain of Klebsiella pneumoniae with Reduced Susceptibility to Expanded-Spectrum Cephalosporins. J. Clin. Microbiol. 42: 4242-4249 [Abstract] [Full Text]  
  • Noren, T., Akerlund, T., Back, E., Sjoberg, L., Persson, I., Alriksson, I., Burman, L. G. (2004). Molecular Epidemiology of Hospital-Associated and Community-Acquired Clostridium difficile Infection in a Swedish County. J. Clin. Microbiol. 42: 3635-3643 [Abstract] [Full Text]  
  • 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 [Abstract] [Full Text]  
  • Svenungsson, B., Burman, L. G., Jalakas-Pornull, K., Lagergren, A., Struwe, J., Akerlund, T. (2003). Epidemiology and Molecular Characterization of Clostridium difficile Strains from Patients with Diarrhea: Low Disease Incidence and Evidence of Limited Cross-Infection in a Swedish Teaching Hospital. J. Clin. Microbiol. 41: 4031-4037 [Abstract] [Full Text]  
  • 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. Bacteriol. 185: 4461-4470 [Abstract] [Full Text]  
  • Schalch, B., Bader, L., Schau, H.-P., Bergmann, R., Rometsch, A., Maydl, G., Kessler, S. (2003). Molecular Typing of Clostridium perfringens from a Food-Borne Disease Outbreak in a Nursing Home: Ribotyping versus Pulsed-Field Gel Electrophoresis. J. Clin. Microbiol. 41: 892-895 [Abstract] [Full Text]  
  • Dridi, L., Tankovic, J., Burghoffer, B., Barbut, F., Petit, J.-C. (2002). gyrA and gyrB Mutations Are Implicated in Cross-Resistance to Ciprofloxacin and Moxifloxacin in Clostridium difficile. Antimicrob. Agents Chemother. 46: 3418-3421 [Abstract] [Full Text]  
  • Klaassen, C. H. W., van Haren, H. A., Horrevorts, A. M. (2002). Molecular Fingerprinting of Clostridium difficile Isolates: Pulsed-Field Gel Electrophoresis versus Amplified Fragment Length Polymorphism. J. Clin. Microbiol. 40: 101-104 [Abstract] [Full Text]  
  • Wong, H.-C., Lin, C.-H. (2001). Evaluation of Typing of Vibrioparahaemolyticus by Three PCR Methods Using Specific Primers. J. Clin. Microbiol. 39: 4233-4240 [Abstract] [Full Text]  
  • KATO, H., KITA, H., KARASAWA, T., MAEGAWA, T., KOINO, Y., TAKAKUWA, H., SAIKAI, T., KOBAYASHI, K., YAMAGISHI, T., NAKAMURA, S. (2001). Colonisation and transmission of Clostridium difficile in healthy individuals examined by PCR ribotyping and pulsed-field gel electrophoresis. J Med Microbiol 50: 720-727 [Abstract] [Full Text]  
  • Kato, H., Kato, N., Watanabe, K., Yamamoto, T., Suzuki, K., Ishigo, S., Kunihiro, S., Nakamura, I., Killgore, G. E., Nakamura, S. (2001). Analysis of Clostridium difficile Isolates from Nosocomial Outbreaks at Three Hospitals in Diverse Areas of Japan. J. Clin. Microbiol. 39: 1391-1395 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»