Summary
Escherichia coli O157:H7 has been recognized since 1982 as a serious human pathogen spread by contaminated food and water. Pulsed-field gel electrophoresis has proven useful for identification of specific isolates/strains of this organism. Hemolytic uremic syndrome (HUS), generally occurring in children or the aged, is the most severe sequela associated with E. coli O157:H7 infection. The presently described work was designed to compare the genomic profile of isolates known to have caused HUS with those having had no such involvement. We asked the question: “Can we develop the means to recognize an ”HUS-prone“ E. coli isolate and thereby alert medical personnel to the increased risk?” Twenty-two HUSrelated and 10 HUS-unrelated E. coli O157:H7 samples were chosen for genomic analysis. Isolates were cultured overnight prior to being embedded in agarose gel plugs. Plugs were digested, using XbaI restriction endonuclease, and subjected to pulsed-field gel electrophoresis (PFGE) for 20 hours. Gels were stained with ethidium bromide, photographed under ultraviolet light, and Southern blotted. Radiolabeled toxin gene probes were used for hybridization assays. The two classes of isolates were compared by optical imaging software. A computer-generated dendrogram, based on restriction profiles, offered strong initial evidence that the HUS sequela may be produced by a particularly virulent and identifiable clone. The predictive value of this finding appears to be substantial.
Keywords
- Hemolytic Uremic Syndrome
- Restriction Profile
- Restriction Fragment Length Polymorphism Pattern
- Ethidium Bromide Solution
- Severe Sequela
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsPreview
Unable to display preview. Download preview PDF.
References
Bohm, H., and H. Karch. 1992. DNA fingerprinting of Escherichia coli 0157:H7 strains by pulsed-field gel electrophoresis. J. Clin. Microbiol. 30: 2169–72.
Carson, C.A., J.M. Keller, K.K. McAdoo, D. Wang, B. Higgins, C.W. Bailey, J.G. Thorne, B.J. Payne, M. Skala, and A.W. Hahn. 1995. Escherichia coli O157:H7 restriction pattern recognition by artificial neural network. J. Clin. Microbiol. 33(11):2894–98.
Chu, G., D. Vollrath, and R.W. Davis. 1986. Separation of large DNA molecules by contour-clamped homogeneous electric field. Science 234: 1582–85.
Harsano, K.D., C.W. Kaspar, and J.B. Luchansky. 1993. Comparison and genomic sizing of Escherichia coli O157:H7 isolates by pulsed-field gel electrophoresis. App. Environ. Microbiol. 59: 3141–44.
Karmali, M. 1989. Infection by verocytotoxin producing Escherichia coli. Clin. Microbiol. Rev. 2 (1): 15–38.
Maniatis, T., et al. Molecular Cloning: A Laboratory Manual. 2nd Edition. Cold Spring Harbor Laboratory Press. 1989.
Ott, M., L. Bender, G. Blum, M. Schmittroth, M. Achtman, H. Tschape, and J. Hacker. 1991. Virulence patterns and long-range genetic mapping of extraintestinal Escherichia coli K I, K5 and KI00 isolates: use of pulsed-field gel electrophoresis. Infect. Immun. 59: 2664–72.
Samadpour M., J. Liston, J. Ongerth, and P. Tarr. 1990. Evaluation of DNA probes for detection of shiga-liketoxin-producing E. coli in food and calf fecal samples. Applied Envir. Microbio1. 56 (5): 1212–15.
Samadpour, M., L.M. Grimm, B. Desai, D. Alfi. J.E. Ongerth, and P.I. Tarr. 1993. Molecular epidemiology of E. 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. 31 (12): 3179–83.
Whittam, T.S., M.L. Wolfe, I.K. Wachsmuth, F. Orskov, I. Orskov, and R.A. Wilson. 1993. Clonal relationships among Escherichia coli strains that cause hemorrhagic colitis and infantile diarrhea. Infect. Immun. 61: 1619–29.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1997 Springer Science+Business Media New York
About this chapter
Cite this chapter
McAdoo, K.K., Carson, C.A. (1997). Evaluation of DNA “Fingerprinting” for Predicting the Potential of E. coli O157:H7 Isolates to Cause Hemolytic Uremic Syndrome (HUS). In: Paul, P.S., Francis, D.H., Benfield, D.A. (eds) Mechanisms in the Pathogenesis of Enteric Diseases. Advances in Experimental Medicine and Biology, vol 412. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-1828-4_28
Download citation
DOI: https://doi.org/10.1007/978-1-4899-1828-4_28
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4899-1830-7
Online ISBN: 978-1-4899-1828-4
eBook Packages: Springer Book Archive