Detection of Escherichia coli O157:H7 by Immunomagnetic Separation and Multiplex Polymerase Chain Reaction

  • Ian G. Wilson
Part of the Methods in Biotechnology book series (MIBT, volume 14)


Since 1983, when Escherichia coli O157:H7 was first recognised as a cause of hemorrhagic colitis and hemolytic uremic syndrome (HUS), verotoxin-producing strains of E. coli (VTEC) have been identified as the cause of increasing numbers of cases of serious human illness (1,2). One of a number of large outbreaks in Scotland resulted in 20 deaths, and a massive outbreak in Japan affected more than 9000 people (3,4). In England and Wales, 90 % of VTEC cases are sporadic (5).


  1. 1.
    Ammon A. (1997) Surveillance of enterohaemorrhagic E. coli (EHEC) infections and haemolytic uraemic syndrome (HUS) in Europe. Euro surveillance 2, 91–96.PubMedGoogle Scholar
  2. 2.
    Armstrong G. L., Hollingsworth J., and Morris J. G. (1986) Emerging foodborne pathogens: Escherichia coli O157:H7 as a model of entry of a new pathogen into the food supply of the developed world. Epidemiol. Rev. 18, 29–51.Google Scholar
  3. 3.
    Anonymous (1996) Enterohaemorrhagic Escherichia coli infection. Weekly Epidemiol. Record 30, 229–230.Google Scholar
  4. 4.
    Gutierrez E. and Netley G. (1997) Japanese Escherichia coli outbreak is still puzzling health officials. Lancet 348, 540.CrossRefGoogle Scholar
  5. 5.
    Parry S. M., Salmon R. L., Willshaw G. A. and Cheasty T. (1998) Risk factors for and prevention of sporadic infections with vero cytotoxin (shiga toxin) producing Escherichia coli O157. Lancet 351, 1019–1022.PubMedCrossRefGoogle Scholar
  6. 6.
    Law D. (1997) The significance of verocytotoxin-producing Escherichia coli other than E. coli O157. PHLS Microbiol. Dig. 14, 72–75.Google Scholar
  7. 7.
    Tarr P. (1994) Escherichia coli O157:H7: overview of clinical and epidemiological issues. J. FoodProt. 57, 632–636.Google Scholar
  8. 8.
    MacDonald I. A. R., Gould I. M. and Curnow J. (1996) Epidemiology of infection due to Escherichia coli O157: a 3-year prospective study. Epidemiol. Infect. 116, 279–284.PubMedCrossRefGoogle Scholar
  9. 9.
    Gill C. O., McGinnis J. C., Rahn K. and Houde A. (1996) The hygienic condition of manufacturing beef destined for the manufacture of hamburger patties. Food Microbiol. 13, 391–396.CrossRefGoogle Scholar
  10. 10.
    Willshaw G. A., Thirlwell J., Jones A. P., et al. (1994) Vero cytotoxin-producing Escherichia coli O157 in beefburgers linked to an outbreak of diarrhoea, haemorrhagic colitis and haemolytic uraemic syndrome in Britain. Lett. Appl. Microbiol. 19, 304–307.PubMedCrossRefGoogle Scholar
  11. 11.
    Besser R. E, Lett S. M., Weber J. T., Doyle M. P., Barrett T., Wells J. G. et al. (1993) An outbreak of diarrhoea and haemolytic uremic syndrome from Escherichia coli O157 in fresh-pressed apple juice. JAMA 111, 2217–2220.CrossRefGoogle Scholar
  12. 12.
    Anonymous (1997) Outbreaks of Escherichia coli O157:H7 infection and cryptosporidiosis associated with drinking unpasteurized apple cider-Connecticut and New York, October 1996. JAMA 277, 781–782.Google Scholar
  13. 13.
    Anonymous (1995) Escherichia coli O157:H7 outbreak linked to commercially distributed dry-cure salami. M.M.W.R. 44(9), 157–160.Google Scholar
  14. 14.
    Sharp J. C. M., Reilly W. J., Coia J. E., Curnow J., and Synge B. A. (1995) Escherichia coli O157 infection in Scotland: an epidemiological overview, 1984-94 PHLS Microbiol. Dig. 12, 134–140.Google Scholar
  15. 15.
    Duncan S. E. and Hackney C. R. (1994) Relevance of Escherichia coli O157:H7 to the dairy industry. Dairy Food Environ. Sanit. 14, 656–660.Google Scholar
  16. 16.
    Morgan D., Newman C. P., Hutchinson D. N., Walker A. M., Rowe B. and Majid F. (1993) Verocytotoxin producing Escherichia coli O157 infections associated with the consumption of yoghurt. Epidemiol. Infect. 111, 181–187.PubMedCrossRefGoogle Scholar
  17. 17.
    Ackman D., Marks S., Mack P., Caldwell M., Root T., and Birkhead G. (1997) Swimming-associated haemorrhagic colitis due to Escherichia coli O157:H7 infection: evidence of prolonged contamination of a fresh water lake. Epidemiol. Infect. 119, 1–8.PubMedCrossRefGoogle Scholar
  18. 18.
    Keene W. E., McAnulty J. M., Hoesly F. C., Williams L. P., Hedberg K., Oxman G. L. et al. (1994) A swimming-associated outbreak of hemorrhagic colitis caused by Escherichia coli O157:H7 and Shigella sonnei. New Engl. J. Med. 331, 579–584.CrossRefGoogle Scholar
  19. 19.
    Anonymous (1997) Outbreaks of Escherichia coli O157:H7 infection associated with eating alfalfa sprouts-Michigan and Virginia, June-July 1997. JAMA 278, 809–810.Google Scholar
  20. 20.
    Bolton F. J., Crozier L., and Williamson J. K. (1996) Isolation of Escherichia coli O157 from raw meat products. Lett. Appl. Microbiol. 23, 317–321.PubMedCrossRefGoogle Scholar
  21. 21.
    Wallace J. S., Cheasty T. and Jones K. (1997) Isolation of verocytotoxin-producing Escherichia coli O157 from wild birds. J. Appl. Microbiol. 82, 399–404.PubMedCrossRefGoogle Scholar
  22. 22.
    Wang G., Zhao T., and Doyle M. P. (1996) Fate of enterohaemorrhagic Escherichia coli O157:H7 in bovine feces. Appl. Environ. Microbiol. 62, 2567–2570.PubMedGoogle Scholar
  23. 23.
    Anonymous (1998) Biomagnetic Techniques in Molecular Biology, 3rd ed. Dynal Oslo, Norway.Google Scholar
  24. 24.
    Safarik I., Safarkov M., and Forsythe S. J. (1995) The application of magnetic separations in applied microbiology. J. Appl. Bacteriol. 78, 575–585.PubMedGoogle Scholar
  25. 25.
    Tomoyasu T. (1998) Improvement of the immunomagnetic separation method selective for Escherichia coli O157 strains. Appl. Environ. Microbiol. 64, 376–382.PubMedGoogle Scholar
  26. 26.
    Chen J., Johnson R., and Griffiths M. (1998) Detection of verotoxigenic Escherichia coli by magnetic capture-hybridization PCR. Appl. Environ. Microbiol. 64, 147–152.PubMedGoogle Scholar
  27. 27.
    Heuvelink A. E., Van de Kar N. C. A. J., Meis J. F. G. M., Monnens L. A. H., and Melchers W. J. G. (1995) Characterization of verocytotoxin-producing Escherichia coli 157 isolated from patients with haemolytic uremic syndrome in Western Europe. Epidemiol. Infect. 115, 1–14.PubMedCrossRefGoogle Scholar
  28. 28.
    Meng J., Zhao S., Doyle M. P., Mitchell S. E., and Kresovich S. (1997) A multiplex PCR for identifying Shiga-like toxin-producing Escherichia coli O157:H7. Lett. Appl. Microbiol. 24, 172–176.PubMedCrossRefGoogle Scholar
  29. 29.
    Roberts D., Hooper W. L., and Greenwood M. (1995) Section 6: Isolation and enrichment of micro-organisms, in Practical Food Microbiology, Methods for the Examination of Food for Organisms of Public Health Significance, PHLS London.Google Scholar
  30. 30.
    Gannon V., King R., Kim J., and Golsteyn Thomas E. (1992) Rapid and sensitive method for detection of Shiga-like toxin-producing Escherichia coli in ground beef using the polymerase chain reaction. Appl. Environ. Microbiol. 58, 3809–3815.PubMedGoogle Scholar
  31. 31.
    Beebakhee G., Louie M., de Azavedo J., and Brunton J. (1992) Cloning and nucleotide sequence of the eae gene homologue from enterohaemorrhagic Escherichia coli serotype O157:H7. FEMS Microbiol. Lett. 91, 63–68.CrossRefGoogle Scholar
  32. 32.
    Widjojoatmodjo M., Fluit A., and Verhoef J. (1994) Rapid identification of bacteria by PCR-single-strand conformation polymorphism. J. Clin. Microbiol. 32, 3002–3007.PubMedGoogle Scholar
  33. 33.
    Bolton E.J., Crozier L., and Williamson J. K. (1995) New technical approaches to Escherichia coli O157. PHLS Microbiol. Digest 12, 67–70.Google Scholar
  34. 34.
    Wilson I. G. (1997) Inhibition and facilitation of nucleic acid amplification. Appl. Environ. Microbiol. 63, 3741–3751.PubMedGoogle Scholar
  35. 35.
    Karch H., Meyer T., Russmann H., and Heeseman J. (1992) Frequent loss of shiga-like toxin genes in clinical isolates of Escherichia coli upon subcultivation. Infect. Immunol. 60, 3464–3467.Google Scholar
  36. 36.
    Widjojoatmodjo M. N., Fluit A. C., Torensma R., Verdonk G. P. and Verhoef J. (1992) The magnetic immuno polymerase chain reaction for direct detection of salmonella in fecal samples. J. Clin. Microbiol. 30, 3195–3199.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2001

Authors and Affiliations

  • Ian G. Wilson
    • 1
  1. 1.Bacteriology DepartmentBelfast City Hospital,BelfastNorthern Ireland

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