Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Jun;33(6):1534–1536. doi: 10.1128/jcm.33.6.1534-1536.1995

Arbitrarily primed PCR DNA fingerprinting of Escherichia coli O157:H7 strains by using templates from boiled cultures.

G Madico 1, N S Akopyants 1, D E Berg 1
PMCID: PMC228210  PMID: 7650181

Abstract

Arbitrarily primed PCR allows genetically different bacterial strains to be distinguished with great sensitivity and efficiency. We report that informative, reproducible arbitrarily primed PCR profiles can be obtained from Escherichia coli O157:H7 strains by using boiled stationary-phase cultures, without the need for time-consuming phenol extraction. This simple template preparation procedure should be especially useful in large epidemiologic studies when many strains must be typed.

Full Text

The Full Text of this article is available as a PDF (431.8 KB).

Selected References

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

  1. Akopyanz N., Bukanov N. O., Westblom T. U., Kresovich S., Berg D. E. DNA diversity among clinical isolates of Helicobacter pylori detected by PCR-based RAPD fingerprinting. Nucleic Acids Res. 1992 Oct 11;20(19):5137–5142. doi: 10.1093/nar/20.19.5137. [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. Chapman P. A., Wright D. J., Siddons C. A. A comparison of immunomagnetic separation and direct culture for the isolation of verocytotoxin-producing Escherichia coli O157 from bovine faeces. J Med Microbiol. 1994 Jun;40(6):424–427. doi: 10.1099/00222615-40-6-424. [DOI] [PubMed] [Google Scholar]
  4. Elaichouni A., van Emmelo J., Claeys G., Verschraegen G., Verhelst R., Vaneechoutte M. Study of the influence of plasmids on the arbitrary primer polymerase chain reaction fingerprint of Escherichia coli strains. FEMS Microbiol Lett. 1994 Jan 15;115(2-3):335–339. doi: 10.1111/j.1574-6968.1994.tb06660.x. [DOI] [PubMed] [Google Scholar]
  5. Griffin P. M., Tauxe R. V. The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. Epidemiol Rev. 1991;13:60–98. doi: 10.1093/oxfordjournals.epirev.a036079. [DOI] [PubMed] [Google Scholar]
  6. MacDonald K. L., Osterholm M. T. The emergence of Escherichia coli O157:H7 infection in the United States. The changing epidemiology of foodborne disease. JAMA. 1993 May 5;269(17):2264–2266. [PubMed] [Google Scholar]
  7. Olsvik O., Popovic T., Skjerve E., Cudjoe K. S., Hornes E., Ugelstad J., Uhlén M. Magnetic separation techniques in diagnostic microbiology. Clin Microbiol Rev. 1994 Jan;7(1):43–54. doi: 10.1128/cmr.7.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Samadpour M., Grimm L. M., Desai B., Alfi D., Ongerth J. E., Tarr P. I. Molecular epidemiology of Escherichia coli O157:H7 strains by bacteriophage lambda restriction fragment length polymorphism analysis: application to a multistate foodborne outbreak and a day-care center cluster. J Clin Microbiol. 1993 Dec;31(12):3179–3183. doi: 10.1128/jcm.31.12.3179-3183.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Wang G., Whittam T. S., Berg C. M., Berg D. E. RAPD (arbitrary primer) PCR is more sensitive than multilocus enzyme electrophoresis for distinguishing related bacterial strains. Nucleic Acids Res. 1993 Dec 25;21(25):5930–5933. doi: 10.1093/nar/21.25.5930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Welsh J., McClelland M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res. 1990 Dec 25;18(24):7213–7218. doi: 10.1093/nar/18.24.7213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Whittam T. S., Wolfe M. L., Wachsmuth I. K., Orskov F., Orskov I., Wilson R. A. Clonal relationships among Escherichia coli strains that cause hemorrhagic colitis and infantile diarrhea. Infect Immun. 1993 May;61(5):1619–1629. doi: 10.1128/iai.61.5.1619-1629.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Williams J. G., Hanafey M. K., Rafalski J. A., Tingey S. V. Genetic analysis using random amplified polymorphic DNA markers. Methods Enzymol. 1993;218:704–740. doi: 10.1016/0076-6879(93)18053-f. [DOI] [PubMed] [Google Scholar]
  13. Williams J. G., Kubelik A. R., Livak K. J., Rafalski J. A., Tingey S. V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990 Nov 25;18(22):6531–6535. doi: 10.1093/nar/18.22.6531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Wright D. J., Chapman P. A., Siddons C. A. Immunomagnetic separation as a sensitive method for isolating Escherichia coli O157 from food samples. Epidemiol Infect. 1994 Aug;113(1):31–39. doi: 10.1017/s0950268800051438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. van Belkum A. DNA fingerprinting of medically important microorganisms by use of PCR. Clin Microbiol Rev. 1994 Apr;7(2):174–184. doi: 10.1128/cmr.7.2.174. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES