Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Aug;33(8):2022–2026. doi: 10.1128/jcm.33.8.2022-2026.1995

In vivo stability and discriminatory power of methicillin-resistant Staphylococcus aureus typing by restriction endonuclease analysis of plasmid DNA compared with those of other molecular methods.

A I Hartstein 1, C L Phelps 1, R Y Kwok 1, M E Mulligan 1
PMCID: PMC228328  PMID: 7559941

Abstract

We evaluated test discriminatory power and DNA type alterations among methicillin-resistant Staphylococcus aureus strains by testing 199 sequential isolates from 39 patients collected over 30 to 228 days. Isolates were typed by one or three different methods (restriction endonuclease analysis of plasmid DNA [REAP] with or without pulsed-field gel electrophoresis of genomic DNA [PFGE] and immunoblotting [IB]). REAP was highly discriminatory compared with PFGE and IB. However, the initial isolates from 4 of the 39 patients lacked detectable plasmid DNA and could not be typed by REAP. Typing of individual patient isolates showed that a different REAP type was identified only once every 138 days. Among 25 comparisons, seven sequential isolate pairs demonstrating REAP differences were also different by PFGE and IB. This likely represented the presence of more than one strain. Eighteen other pairs with REAP differences were identical or related to one another by PFGE and IB typing, and 17 of these differences were likely caused by a single genetic alteration within the same strain or clone. The rate of PFGE differences explicable by single genetic alterations among sequential isolates identical by REAP was similar to the overall rate for REAP differences in the whole collection. We conclude that REAP and PFGE typing differences explicable by single genetic alterations are relatively infrequent but not rare. These isolates should be examined by alternative typing systems to further support or refute clonality.

Full Text

The Full Text of this article is available as a PDF (684.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arbeit R. D., Arthur M., Dunn R., Kim C., Selander R. K., Goldstein R. Resolution of recent evolutionary divergence among Escherichia coli from related lineages: the application of pulsed field electrophoresis to molecular epidemiology. J Infect Dis. 1990 Feb;161(2):230–235. doi: 10.1093/infdis/161.2.230. [DOI] [PubMed] [Google Scholar]
  2. Boyce J. M., Jackson M. M., Pugliese G., Batt M. D., Fleming D., Garner J. S., Hartstein A. I., Kauffman C. A., Simmons M., Weinstein R. Methicillin-resistant Staphylococcus aureus (MRSA): a briefing for acute care hospitals and nursing facilities. The AHA Technical Panel on Infections Within Hospitals. Infect Control Hosp Epidemiol. 1994 Feb;15(2):105–115. doi: 10.1086/646870. [DOI] [PubMed] [Google Scholar]
  3. Chetchotisakd P., Phelps C. L., Hartstein A. I. Assessment of bacterial cross-transmission as a cause of infections in patients in intensive care units. Clin Infect Dis. 1994 Jun;18(6):929–937. doi: 10.1093/clinids/18.6.929. [DOI] [PubMed] [Google Scholar]
  4. Doebbeling B. N., Pfaller M. A., Hollis R. J., Boyken L. D., Pignatari A. C., Herwaldt L. A., Wenzel R. P. Restriction endonuclease analysis of Staphylococcus aureus plasmid DNA from three continents. Eur J Clin Microbiol Infect Dis. 1992 Jan;11(1):4–8. doi: 10.1007/BF01971263. [DOI] [PubMed] [Google Scholar]
  5. Goering R. V. Molecular epidemiology of nosocomial infection: analysis of chromosomal restriction fragment patterns by pulsed-field gel electrophoresis. Infect Control Hosp Epidemiol. 1993 Oct;14(10):595–600. doi: 10.1086/646645. [DOI] [PubMed] [Google Scholar]
  6. Hall L. M. Are point mutations or DNA rearrangements responsible for the restriction fragment length polymorphisms that are used to type bacteria? Microbiology. 1994 Jan;140(Pt 1):197–204. doi: 10.1099/13500872-140-1-197. [DOI] [PubMed] [Google Scholar]
  7. Hartstein A. I., Morthland V. H., Eng S., Archer G. L., Schoenknecht F. D., Rashad A. L. Restriction enzyme analysis of plasmid DNA and bacteriophage typing of paired Staphylococcus aureus blood culture isolates. J Clin Microbiol. 1989 Aug;27(8):1874–1879. doi: 10.1128/jcm.27.8.1874-1879.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hartstein A. I., Mulligan M. E., Morthland V. H., Kwok R. Y. Recurrent Staphylococcus aureus bacteremia. J Clin Microbiol. 1992 Mar;30(3):670–674. doi: 10.1128/jcm.30.3.670-674.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kreiswirth B., Kornblum J., Arbeit R. D., Eisner W., Maslow J. N., McGeer A., Low D. E., Novick R. P. Evidence for a clonal origin of methicillin resistance in Staphylococcus aureus. Science. 1993 Jan 8;259(5092):227–230. doi: 10.1126/science.8093647. [DOI] [PubMed] [Google Scholar]
  10. Layton M. C., Hierholzer W. J., Jr, Patterson J. E. The evolving epidemiology of methicillin-resistant Staphylococcus aureus at a university hospital. Infect Control Hosp Epidemiol. 1995 Jan;16(1):12–17. doi: 10.1086/646996. [DOI] [PubMed] [Google Scholar]
  11. Maslow J. N., Mulligan M. E., Arbeit R. D. Molecular epidemiology: application of contemporary techniques to the typing of microorganisms. Clin Infect Dis. 1993 Aug;17(2):153–164. doi: 10.1093/clinids/17.2.153. [DOI] [PubMed] [Google Scholar]
  12. Mulligan M. E., Arbeit R. D. Epidemiologic and clinical utility of typing systems for differentiating among strains of methicillin-resistant Staphylococcus aureus. Infect Control Hosp Epidemiol. 1991 Jan;12(1):20–28. doi: 10.1086/646234. [DOI] [PubMed] [Google Scholar]
  13. Mulligan M. E., Kwok R. Y., Citron D. M., John J. F., Jr, Smith P. B. Immunoblots, antimicrobial resistance, and bacteriophage typing of oxacillin-resistant Staphylococcus aureus. J Clin Microbiol. 1988 Nov;26(11):2395–2401. doi: 10.1128/jcm.26.11.2395-2401.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mulligan M. E., Murray-Leisure K. A., Ribner B. S., Standiford H. C., John J. F., Korvick J. A., Kauffman C. A., Yu V. L. Methicillin-resistant Staphylococcus aureus: a consensus review of the microbiology, pathogenesis, and epidemiology with implications for prevention and management. Am J Med. 1993 Mar;94(3):313–328. doi: 10.1016/0002-9343(93)90063-u. [DOI] [PubMed] [Google Scholar]
  15. Smith C. L., Cantor C. R. Purification, specific fragmentation, and separation of large DNA molecules. Methods Enzymol. 1987;155:449–467. doi: 10.1016/0076-6879(87)55030-3. [DOI] [PubMed] [Google Scholar]
  16. Tenover F. C., Arbeit R., Archer G., Biddle J., Byrne S., Goering R., Hancock G., Hébert G. A., Hill B., Hollis R. Comparison of traditional and molecular methods of typing isolates of Staphylococcus aureus. J Clin Microbiol. 1994 Feb;32(2):407–415. doi: 10.1128/jcm.32.2.407-415.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Trilla A., Nettleman M. D., Hollis R. J., Fredrickson M., Wenzel R. P., Pfaller M. A. Restriction endonuclease analysis of plasmid DNA from methicillin-resistant Staphylococcus aureus: clinical application over a three-year period. Infect Control Hosp Epidemiol. 1993 Jan;14(1):29–35. doi: 10.1086/646627. [DOI] [PubMed] [Google Scholar]
  18. Zierdt C. H. Long-term Staphylococcus aureus carrier state in hospital patients. J Clin Microbiol. 1982 Sep;16(3):517–520. doi: 10.1128/jcm.16.3.517-520.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES