Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Nov;35(11):2977–2980. doi: 10.1128/jcm.35.11.2977-2980.1997

Rapid pulsed-field gel electrophoresis protocol for typing of Escherichia coli O157:H7 and other gram-negative organisms in 1 day.

R K Gautom 1
PMCID: PMC230100  PMID: 9350772

Abstract

Genomic DNA patterns generated by pulsed-field gel electrophoresis are highly specific for different strains of an organism and have significant value in epidemiologic investigations of infectious-disease outbreaks. Unfortunately, time-consuming and tedious specimen processing is an inherent problem which limits the use of this powerful technology as a real-time epidemic investigational tool. Here, I describe a rapid method to improve the response time and provide specific bacterial strain identification for the typing of Escherichia coli O157:H7 and other gram-negative organisms in a single day.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

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

  1. Bannerman T. L., Hancock G. A., Tenover F. C., Miller J. M. Pulsed-field gel electrophoresis as a replacement for bacteriophage typing of Staphylococcus aureus. J Clin Microbiol. 1995 Mar;33(3):551–555. doi: 10.1128/jcm.33.3.551-555.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barrett T. J., Lior H., Green J. H., Khakhria R., Wells J. G., Bell B. P., Greene K. D., Lewis J., Griffin P. M. Laboratory investigation of a multistate food-borne outbreak of Escherichia coli O157:H7 by using pulsed-field gel electrophoresis and phage typing. J Clin Microbiol. 1994 Dec;32(12):3013–3017. doi: 10.1128/jcm.32.12.3013-3017.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brosch R., Buchrieser C., Rocourt J. Subtyping of Listeria monocytogenes serovar 4b by use of low-frequency-cleavage restriction endonucleases and pulsed-field gel electrophoresis. Res Microbiol. 1991 Jul-Aug;142(6):667–675. doi: 10.1016/0923-2508(91)90080-t. [DOI] [PubMed] [Google Scholar]
  4. Böhm H., Karch H. DNA fingerprinting of Escherichia coli O157:H7 strains by pulsed-field gel electrophoresis. J Clin Microbiol. 1992 Aug;30(8):2169–2172. doi: 10.1128/jcm.30.8.2169-2172.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cameron D. N., Khambaty F. M., Wachsmuth I. K., Tauxe R. V., Barrett T. J. Molecular characterization of Vibrio cholerae O1 strains by pulsed-field gel electrophoresis. J Clin Microbiol. 1994 Jul;32(7):1685–1690. doi: 10.1128/jcm.32.7.1685-1690.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goering R. V., Winters M. A. Rapid method for epidemiological evaluation of gram-positive cocci by field inversion gel electrophoresis. J Clin Microbiol. 1992 Mar;30(3):577–580. doi: 10.1128/jcm.30.3.577-580.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jackson L. A., Alexander E. R., DeBolt C. A., Swenson P. D., Boase J., McDowell M. G., Reeves M. W., Wenger J. D. Evaluation of the use of mass chemoprophylaxis during a school outbreak of enzyme type 5 serogroup B meningococcal disease. Pediatr Infect Dis J. 1996 Nov;15(11):992–998. doi: 10.1097/00006454-199611000-00012. [DOI] [PubMed] [Google Scholar]
  8. Johnson W. M., Bernard K., Marrie T. J., Tyler S. D. Discriminatory genomic fingerprinting of Legionella pneumophila by pulsed-field electrophoresis. J Clin Microbiol. 1994 Oct;32(10):2620–2621. doi: 10.1128/jcm.32.10.2620-2621.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kersulyte D., Struelens M. J., Deplano A., Berg D. E. Comparison of arbitrarily primed PCR and macrorestriction (pulsed-field gel electrophoresis) typing of Pseudomonas aeruginosa strains from cystic fibrosis patients. J Clin Microbiol. 1995 Aug;33(8):2216–2219. doi: 10.1128/jcm.33.8.2216-2219.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Leonard R. B., Mayer J., Sasser M., Woods M. L., Mooney B. R., Brinton B. G., Newcomb-Gayman P. L., Carroll K. C. Comparison of MIDI Sherlock system and pulsed-field gel electrophoresis in characterizing strains of methicillin-resistant Staphylococcus aureus from a recent hospital outbreak. J Clin Microbiol. 1995 Oct;33(10):2723–2727. doi: 10.1128/jcm.33.10.2723-2727.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Matushek M. G., Bonten M. J., Hayden M. K. Rapid preparation of bacterial DNA for pulsed-field gel electrophoresis. J Clin Microbiol. 1996 Oct;34(10):2598–2600. doi: 10.1128/jcm.34.10.2598-2600.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Schlichting C., Branger C., Fournier J. M., Witte W., Boutonnier A., Wolz C., Goullet P., Döring G. Typing of Staphylococcus aureus by pulsed-field gel electrophoresis, zymotyping, capsular typing, and phage typing: resolution of clonal relationships. J Clin Microbiol. 1993 Feb;31(2):227–232. doi: 10.1128/jcm.31.2.227-232.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schoonmaker D., Heimberger T., Birkhead G. Comparison of ribotyping and restriction enzyme analysis using pulsed-field gel electrophoresis for distinguishing Legionella pneumophila isolates obtained during a nosocomial outbreak. J Clin Microbiol. 1992 Jun;30(6):1491–1498. doi: 10.1128/jcm.30.6.1491-1498.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Tenover F. C., Arbeit R. D., Goering R. V., Mickelsen P. A., Murray B. E., Persing D. H., Swaminathan B. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol. 1995 Sep;33(9):2233–2239. doi: 10.1128/jcm.33.9.2233-2239.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Yakubu D. E., Abadi F. J., Pennington T. H. Molecular epidemiology of recent United Kingdom isolates of Neisseria meningitidis serogroup C. Epidemiol Infect. 1994 Aug;113(1):53–65. doi: 10.1017/s0950268800051463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Yakubu D. E., Pennington T. H. Epidemiological evaluation of Neisseria meningitidis serogroup B by pulsed-field gel electrophoresis. FEMS Immunol Med Microbiol. 1995 Feb;10(3-4):185–189. doi: 10.1111/j.1574-695X.1995.tb00032.x. [DOI] [PubMed] [Google Scholar]
  19. de Moissac Y. R., Ronald S. L., Peppler M. S. Use of pulsed-field gel electrophoresis for epidemiological study of Bordetella pertussis in a whooping cough outbreak. J Clin Microbiol. 1994 Feb;32(2):398–402. doi: 10.1128/jcm.32.2.398-402.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES