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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1994 Sep;32(9):2067–2070. doi: 10.1128/jcm.32.9.2067-2070.1994

Application of typing by pulsed-field gel electrophoresis to the study of Clostridium difficile in a neonatal intensive care unit.

H Kato 1, N Kato 1, K Watanabe 1, K Ueno 1, H Ushijima 1, S Hashira 1, T Abe 1
PMCID: PMC263943  PMID: 7814526

Abstract

Pulsed-field gel electrophoresis (PFGE) analysis of restriction pattern polymorphism was applied to type Clostridium difficile isolated from neonates hospitalized in a neonatal intensive care unit, and the results were compared with those of immunoblot analysis. C. difficile was isolated from fecal specimens of 41 (61%) of 67 neonates during a 5-month investigation. All of these neonates were asymptomatic. Fifty-five C. difficile isolates from 32 patients were analyzed by PFGE after digestion with SmaI and SacII endonucleases and by immunoblotting with 10 different antisera. Fifty-three of 55 isolates from 30 patients were identical by PFGE analysis after SmaI and SacII digestion and immunoblot analysis. Two isolates were different from each other and from the epidemic group by both PFGE and immunoblot analysis. All 53 epidemic isolates were nontoxigenic, while the two remaining isolates were toxigenic. These results suggest that nosocomial spread of nontoxigenic C. difficile infection in the neonatal intensive care unit and suggest that both PFGE and immunoblot are powerful typing tools for the epidemiological study of C. difficile.

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Selected References

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  1. Al-Jumaili I. J., Shibley M., Lishman A. H., Record C. O. Incidence and origin of Clostridium difficile in neonates. J Clin Microbiol. 1984 Jan;19(1):77–78. doi: 10.1128/jcm.19.1.77-78.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allardet-Servent A., Bouziges N., Carles-Nurit M. J., Bourg G., Gouby A., Ramuz M. Use of low-frequency-cleavage restriction endonucleases for DNA analysis in epidemiological investigations of nosocomial bacterial infections. J Clin Microbiol. 1989 Sep;27(9):2057–2061. doi: 10.1128/jcm.27.9.2057-2061.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blaschek H. P., Klacik M. A. Role of DNase in recovery of plasmid DNA from Clostridium perfringens. Appl Environ Microbiol. 1984 Jul;48(1):178–181. doi: 10.1128/aem.48.1.178-181.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bolton R. P., Tait S. K., Dear P. R., Losowsky M. S. Asymptomatic neonatal colonisation by Clostridium difficile. Arch Dis Child. 1984 May;59(5):466–472. doi: 10.1136/adc.59.5.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brefort G., Magot M., Ionesco H., Sebald M. Characterization and transferability of Clostridium perfringens plasmids. Plasmid. 1977 Nov;1(1):52–66. doi: 10.1016/0147-619x(77)90008-7. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Delmée M., Laroche Y., Avesani V., Cornelis G. Comparison of serogrouping and polyacrylamide gel electrophoresis for typing Clostridium difficile. J Clin Microbiol. 1986 Dec;24(6):991–994. doi: 10.1128/jcm.24.6.991-994.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Delmée M., Verellen G., Avesani V., Francois G. Clostridium difficile in neonates: serogrouping and epidemiology. Eur J Pediatr. 1988 Jan;147(1):36–40. doi: 10.1007/BF00442608. [DOI] [PubMed] [Google Scholar]
  9. Devlin H. R., Au W., Foux L., Bradbury W. C. Restriction endonuclease analysis of nosocomial isolates of Clostridium difficile. J Clin Microbiol. 1987 Nov;25(11):2168–2172. doi: 10.1128/jcm.25.11.2168-2172.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kato H., Cavallaro J. J., Kato N., Bartley S. L., Killgore G. E., Watanabe K., Ueno K. Typing of Clostridium difficile by western immunoblotting with 10 different antisera. J Clin Microbiol. 1993 Feb;31(2):413–415. doi: 10.1128/jcm.31.2.413-415.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kato N., Ou C. Y., Kato H., Bartley S. L., Brown V. K., Dowell V. R., Jr, Ueno K. Identification of toxigenic Clostridium difficile by the polymerase chain reaction. J Clin Microbiol. 1991 Jan;29(1):33–37. doi: 10.1128/jcm.29.1.33-37.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Murray B. E., Singh K. V., Heath J. D., Sharma B. R., Weinstock G. M. Comparison of genomic DNAs of different enterococcal isolates using restriction endonucleases with infrequent recognition sites. J Clin Microbiol. 1990 Sep;28(9):2059–2063. doi: 10.1128/jcm.28.9.2059-2063.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Phua T. J., Rogers T. R., Pallett A. P. Prospective study of Clostridium difficile colonization and paracresol detection in the stools of babies on a special care unit. J Hyg (Lond) 1984 Aug;93(1):17–25. doi: 10.1017/s0022172400060885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Simor A. E., Yake S. L., Tsimidis K. Infection due to Clostridium difficile among elderly residents of a long-term-care facility. Clin Infect Dis. 1993 Oct;17(4):672–678. doi: 10.1093/clinids/17.4.672. [DOI] [PubMed] [Google Scholar]
  17. Zedd A. J., Sell T. L., Schaberg D. R., Fekety F. R., Cooperstock M. S. Nosocomial Clostridium difficile reservoir in a neonatal intensive care unit. Pediatr Infect Dis. 1984 Sep-Oct;3(5):429–432. doi: 10.1097/00006454-198409000-00008. [DOI] [PubMed] [Google Scholar]

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