Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1992 Mar;30(3):577–580. doi: 10.1128/jcm.30.3.577-580.1992

Rapid method for epidemiological evaluation of gram-positive cocci by field inversion gel electrophoresis.

R V Goering 1, M A Winters 1
PMCID: PMC265112  PMID: 1313045

Abstract

We report a rapid method for the isolation of intact chromosomal DNA from gram-positive cocci that is suitable for in situ restriction endonuclease digestion in agarose blocks. When combined with a rapid field inversion gel electrophoresis protocol, this approach allows the preparation and electrophoretic analysis of chromosomal restriction fragments produced by rare-cutting enzymes in a total time period of 2 days from start to finish. The utility of the method is demonstrated in the epidemiological evaluation of Staphylococcus epidermidis clusters from two hospitals as well as of additional representative staphylococci and enterococci.

Full text

PDF
577

Images in this article

578Image
on p.578
579Image
on p.579

Selected References

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

  1. Anderson D. J., Kuhns J. S., Vasil M. L., Gerding D. N., Janoff E. N. DNA fingerprinting by pulsed field gel electrophoresis and ribotyping to distinguish Pseudomonas cepacia isolates from a nosocomial outbreak. J Clin Microbiol. 1991 Mar;29(3):648–649. doi: 10.1128/jcm.29.3.648-649.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Carle G. F., Frank M., Olson M. V. Electrophoretic separations of large DNA molecules by periodic inversion of the electric field. Science. 1986 Apr 4;232(4746):65–68. doi: 10.1126/science.3952500. [DOI] [PubMed] [Google Scholar]
  4. Chu G., Vollrath D., Davis R. W. Separation of large DNA molecules by contour-clamped homogeneous electric fields. Science. 1986 Dec 19;234(4783):1582–1585. doi: 10.1126/science.3538420. [DOI] [PubMed] [Google Scholar]
  5. Flanagan J. L., Ventra L., Weiss A. S. Rapid method for preparation and cleavage of bacterial DNA for pulsed-field gel electrophoresis. Nucleic Acids Res. 1989 Jan 25;17(2):814–814. doi: 10.1093/nar/17.2.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fritz R. B., Musich P. R. Unexpected loss of genomic DNA from agarose gel plugs. Biotechniques. 1990 Nov;9(5):542, 544, 546-50. [PubMed] [Google Scholar]
  7. Goering R. V., Bauernfeind A., Lenz W., Przyklenk B. Staphylococcus aureus in patients with cystic fibrosis: an epidemiological analysis using a combination of traditional and molecular methods. Infection. 1990 Jan-Feb;18(1):57–60. doi: 10.1007/BF01644187. [DOI] [PubMed] [Google Scholar]
  8. Goering R. V., Duensing T. D. Rapid field inversion gel electrophoresis in combination with an rRNA gene probe in the epidemiological evaluation of staphylococci. J Clin Microbiol. 1990 Mar;28(3):426–429. doi: 10.1128/jcm.28.3.426-429.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Heinzen R., Stiegler G. L., Whiting L. L., Schmitt S. A., Mallavia L. P., Frazier M. E. Use of pulsed field gel electrophoresis to differentiate Coxiella burnetii strains. Ann N Y Acad Sci. 1990;590:504–513. doi: 10.1111/j.1749-6632.1990.tb42260.x. [DOI] [PubMed] [Google Scholar]
  10. Hébert G. A., Cooksey R. C., Clark N. C., Hill B. C., Jarvis W. R., Thornsberry C. Biotyping coagulase-negative staphylococci. J Clin Microbiol. 1988 Oct;26(10):1950–1956. doi: 10.1128/jcm.26.10.1950-1956.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Le Bourgeois P., Mata M., Ritzenthaler P. Genome comparison of Lactococcus strains by pulsed-field gel electrophoresis. FEMS Microbiol Lett. 1989 May;50(1-2):65–69. doi: 10.1016/0378-1097(89)90460-6. [DOI] [PubMed] [Google Scholar]
  12. Lévy-Frébault V. V., Thorel M. F., Varnerot A., Gicquel B. DNA polymorphism in Mycobacterium paratuberculosis, "wood pigeon mycobacteria," and related mycobacteria analyzed by field inversion gel electrophoresis. J Clin Microbiol. 1989 Dec;27(12):2823–2826. doi: 10.1128/jcm.27.12.2823-2826.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Schwartz D. C., Saffran W., Welsh J., Haas R., Goldenberg M., Cantor C. R. New techniques for purifying large DNAs and studying their properties and packaging. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 1):189–195. doi: 10.1101/sqb.1983.047.01.024. [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. Sobral B. W., Atherly A. G. A rapid and cost-effective method for preparing genomic DNA from gram-negative bacteria in agarose plugs for pulsed-field gel electrophoresis. Biotechniques. 1989 Oct;7(9):938–938. [PubMed] [Google Scholar]

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

RESOURCES