Abstract
From January to March 1993, a suspected outbreak of antibiotic-associated diarrhea occurred on a pediatric oncology ward of the Clinical Center Hospital at the National Institutes of Health. Isolates of Clostridium difficile obtained from six patients implicated in this outbreak were typed by both PCR amplification of rRNA intergenic spacer regions (PCR ribotyping) and restriction endonuclease analysis of genomic DNA. Comparable results were obtained with both methods; five of the six patients were infected with the same strain of C. difficile. Subsequent analysis of 102 C. difficile isolates obtained from symptomatic patients throughout the Clinical Center revealed the existence of 41 distinct and reproducible PCR ribotypes. These data suggest that PCR ribotyping provides a discriminatory, reproducible, and simple alternative to conventional molecular approaches for typing strains of C. difficile.
Full Text
The Full Text of this article is available as a PDF (473.7 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Barbut F., Mario N., Frottier J., Petit J. C. Use of the arbitrary primer polymerase chain reaction for investigating an outbreak of Clostridium difficile-associated diarrhea in AIDS patients. Eur J Clin Microbiol Infect Dis. 1993 Oct;12(10):794–795. doi: 10.1007/BF02098477. [DOI] [PubMed] [Google Scholar]
- Boros I., Kiss A., Venetianer P. Physical map of the seven ribosomal RNA genes of Escherichia coli. Nucleic Acids Res. 1979;6(5):1817–1830. doi: 10.1093/nar/6.5.1817. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Böddinghaus B., Rogall T., Flohr T., Blöcker H., Böttger E. C. Detection and identification of mycobacteria by amplification of rRNA. J Clin Microbiol. 1990 Aug;28(8):1751–1759. doi: 10.1128/jcm.28.8.1751-1759.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cartwright C. P., Stock F., Gill V. J. Improved enrichment broth for cultivation of fastidious organisms. J Clin Microbiol. 1994 Jul;32(7):1825–1826. doi: 10.1128/jcm.32.7.1825-1826.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clabots C. R., Johnson S., Bettin K. M., Mathie P. A., Mulligan M. E., Schaberg D. R., Peterson L. R., Gerding D. N. Development of a rapid and efficient restriction endonuclease analysis typing system for Clostridium difficile and correlation with other typing systems. J Clin Microbiol. 1993 Jul;31(7):1870–1875. doi: 10.1128/jcm.31.7.1870-1875.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clabots C. R., Peterson L. R., Gerding D. N. Characterization of a nosocomial Clostridium difficile outbreak by using plasmid profile typing and clindamycin susceptibility testing. J Infect Dis. 1988 Oct;158(4):731–736. doi: 10.1093/infdis/158.4.731. [DOI] [PubMed] [Google Scholar]
- Delmée M., Avesani V. Correlation between serogroup and susceptibility to chloramphenicol, clindamycin, erythromycin, rifampicin and tetracycline among 308 isolates of Clostridium difficile. J Antimicrob Chemother. 1988 Sep;22(3):325–331. doi: 10.1093/jac/22.3.325. [DOI] [PubMed] [Google Scholar]
- Gürtler V. Typing of Clostridium difficile strains by PCR-amplification of variable length 16S-23S rDNA spacer regions. J Gen Microbiol. 1993 Dec;139(12):3089–3097. doi: 10.1099/00221287-139-12-3089. [DOI] [PubMed] [Google Scholar]
- Heard S. R., Rasburn B., Matthews R. C., Tabaqchali S. Immunoblotting to demonstrate antigenic and immunogenic differences among nine standard strains of Clostridium difficile. J Clin Microbiol. 1986 Sep;24(3):384–387. doi: 10.1128/jcm.24.3.384-387.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Killgore G. E., Kato H. Use of arbitrary primer PCR to type Clostridium difficile and comparison of results with those by immunoblot typing. J Clin Microbiol. 1994 Jun;32(6):1591–1593. doi: 10.1128/jcm.32.6.1591-1593.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kiss A., Sain B., Venetianer P. The number of rRNA genes in Escherichia coli. FEBS Lett. 1977 Jul 1;79(1):77–79. doi: 10.1016/0014-5793(77)80354-2. [DOI] [PubMed] [Google Scholar]
- Kostman J. R., Edlind T. D., LiPuma J. J., Stull T. L. Molecular epidemiology of Pseudomonas cepacia determined by polymerase chain reaction ribotyping. J Clin Microbiol. 1992 Aug;30(8):2084–2087. doi: 10.1128/jcm.30.8.2084-2087.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lambert-Zechovsky N., Bingen E., Denamur E., Brahimi N., Brun P., Mathieu H., Elion J. Molecular analysis provides evidence for the endogenous origin of bacteremia and meningitis due to Enterobacter cloacae in an infant. Clin Infect Dis. 1992 Jul;15(1):30–32. doi: 10.1093/clinids/15.1.30. [DOI] [PubMed] [Google Scholar]
- Lyerly D. M., Krivan H. C., Wilkins T. D. Clostridium difficile: its disease and toxins. Clin Microbiol Rev. 1988 Jan;1(1):1–18. doi: 10.1128/cmr.1.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matar G. M., Swaminathan B., Hunter S. B., Slater L. N., Welch D. F. Polymerase chain reaction-based restriction fragment length polymorphism analysis of a fragment of the ribosomal operon from Rochalimaea species for subtyping. J Clin Microbiol. 1993 Jul;31(7):1730–1734. doi: 10.1128/jcm.31.7.1730-1734.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McFarland L. V., Mulligan M. E., Kwok R. Y., Stamm W. E. Nosocomial acquisition of Clostridium difficile infection. N Engl J Med. 1989 Jan 26;320(4):204–210. doi: 10.1056/NEJM198901263200402. [DOI] [PubMed] [Google Scholar]
- Saulnier P., Bourneix C., Prévost G., Andremont A. Random amplified polymorphic DNA assay is less discriminant than pulsed-field gel electrophoresis for typing strains of methicillin-resistant Staphylococcus aureus. J Clin Microbiol. 1993 Apr;31(4):982–985. doi: 10.1128/jcm.31.4.982-985.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Toma S., Lesiak G., Magus M., Lo H. L., Delmée M. Serotyping of Clostridium difficile. J Clin Microbiol. 1988 Mar;26(3):426–428. doi: 10.1128/jcm.26.3.426-428.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]