Skip to main content Accessibility help

Sequence heterogeneity of the eae gene and detection of verotoxin-producing Escherichia coli using serotype-specific primers

  • M. Louie (a1), J. De Azavedo (a1), R. Clarke (a2), A. Borczyk (a3), H. Lior (a4), M. Richter (a5) and J. Brunton (a1)...


The distribution of the Escherichia coli attaching and effacing (eae) gene in strains of verotoxin-producing E. coli (VTEC) isolated from cattle and humans was studied. The majority of strains isolated from humans with bloody diarrhoea or HUS and cattle with severe diarrhoea were eae. positive (82 and 83% respectively). In contrast. 59% of VTEC isolated from asymptomatic cattle were eae negative and of the remaining 41 % that were eae positive, the majority were serotype O157.H7. The nucleotide sequence of the 3′ end of the eae gene of enteropathogenic E. coli (EPEC) of serotype O55.H7 was found to be almost identical to that of serotype O157. H7. Specific primers are described which detect the eae sequences of VTEC serotypes O157.H7, 0157.H, and EPEC serotypes O55.H7 and O55.H. The nucleotide sequence of the 3′ end of the eae gene of serotype O111.H8 differed significantly from that of O157.H7. Primers were developed to specifically identify the eae sequences of VTEC serotypes O111. H and O111.H8. We conclude that whereas the majority of VTEC associated with disease in cattle and humans possess the eae gene, the gene itself may not be necessary to produce haemorrhagic colitis and HUS. Sequence heterogeneity in the 3′ end of eae alleles of VTEC permits specific identification of subsets of these organisms.

    • Send article to Kindle

      To send this article to your Kindle, first ensure is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about sending to your Kindle. Find out more about sending to your Kindle.

      Note you can select to send to either the or variations. ‘’ emails are free but can only be sent to your device when it is connected to wi-fi. ‘’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

      Find out more about the Kindle Personal Document Service.

      Sequence heterogeneity of the eae gene and detection of verotoxin-producing Escherichia coli using serotype-specific primers
      Available formats

      Send article to Dropbox

      To send this article to your Dropbox account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Dropbox.

      Sequence heterogeneity of the eae gene and detection of verotoxin-producing Escherichia coli using serotype-specific primers
      Available formats

      Send article to Google Drive

      To send this article to your Google Drive account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Google Drive.

      Sequence heterogeneity of the eae gene and detection of verotoxin-producing Escherichia coli using serotype-specific primers
      Available formats


Corresponding author

* Corresponding author: Dr James L. Brunton. The Toronto Hospital. Department of Microbiology. NU 13–124. 200 Elizabeth Street. Toronto. Ontario M5G 2C4.


Hide All
1.Griffin, PM. Ostroff, SM. Tauxe, RV. et al. Illnesses associated with Escherichia coli O157:H7 infections: A broad clinical spectrum. Ann Intern Med 1988; 109: 705–12.
2.Karmali, MA. Infection by verocytotoxin-producing Escherichia coli. Clin Microbiol Rev 1989; 2: 1538.
3.Pai, CH. Ahmed, N, Lior, H. Johnson, WM, Sims, HV. Woods, DE. Epidemiology of sporadic diarrhoea due to verocytotoxin-producing Escherichia coli: A two-year prospective study. J infect Dis 1988; 157: 1054–7.
4.Riley, LW. Remis, RS. Helgerson, SD. et al. Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med 1983; 308: 681–5.
5.Levine, MM. Escherichia coli that cause diarrhoea: Enterotoxigenic, enteropathogenic. enteroinvasive. enterohemorrhagic and enteroadherent. J Infect Dis 1987; 155: 377–89.
6.Belongia, EA. Mac-Donald, KL. Blaser, MJ. Soler, JT. Ammend, DA. Osterholm, MT. Outbreak of Escherichia coli O157: H7 in a day care center: patterns of transmission and control methods. Program Abstr. 1989; 29th Intersci Conf Antimicro Agent Chemother abstr 10.
7.Carter, AO. Borczyk, AA. Carlson, JAK. et al. A severe outbreak of Escherichia coli O157: H7 associated hemorrhagic colitis in a nursing home. N Engl J Med 1987; 317: 1496–500.
8.Ryan, CA. Tauxe, RV. Hosek, GW. et al. Escherichia coli O157: H7 diarrhea in a nursing home: clinical, epidemiological, and pathological findings. J Infect Dis 1986; 154: 631–8.
9.Spika, JS. Parsons, JE. Nordenberg, D. Gunn, RA. Hemolytic uremic syndrome and diarrhea associated with Escherichia coli O157: H7 in a day-care center. J Pediatr 1986; 109: 287–91.
10.Belongia, EA. MacDonald, KL. Parham, GL. et al. An outbreak of Escherichia coli O157: H7 colitis associated with consumption of precooked meat patties. J Infect Dis 1991; 164: 338–43.
11.Clarke, RC. McEwen, SA. Gannon, VP. Lior, H. Gyles, CL. Isolation of verocytotoxin-producing Escherichia coli from milk filters in south-western Ontario. Epidemiol Infect 1989; 102: 253–60.
12.Duncan, L, Mai, V. Carter, A. Carlson, JAK. Borezyk, AA. Karmali, MA. Outbreak of gastrointestinal disease-Ontario. Can Dis Wkly Rep 1987; 13–2: 58.
13.Montenegro, MA. Bulte, M. Trumpf, T. et al. Detection and characterization of fecal verotoxin-producing Escherichia coli from healthy cattle. J Clin Microbiol 1990; 28: 1417–21.
14.Read, SC. Gyles, CL. Clarke, RC. Lior, H, McEwen, S. Prevalence of verocytotoxigenie Escherichia coli in ground beef, pork, and chicken in southwestern Ontario. Epidemiol Infect 1990; 105: 1120.
15.Sherwood, D. Snodgrass, DR. O'Brien, AD. Shiga-like toxin production from Escherichia coli associated with calf diarrhoea. Vet Rec 1985; 116: 217–18.
16.Wells, JG. Shipman, LD. Greene, KD. et al. Isolation of Escherichia coli serotype O157: H7 and other Shiga-like-toxin-producing E. coli from dairy cattle. J Clin Microbiol 1991; 29: 985–9.
17.Wilson, JB. McEwen, SA. Clarke, RC. et al. Distribution and characteristics of verocytotoxigenic Escherichia coli isolated from Ontario dairy cattle. Epidemiol Infect 1992; 108: 423–39.
18.Borczyk, AA. Karmali, MA. Lior, H. Duncan, LMC. Bovine reservoir for verotoxin-producing Escherichia coli O157: H7. Lancet: 1987; i: 98.
19.Orskov, F. Orskov, I. Villar, J. Cattle as a reservoir of verotoxin-producing Escherichia coli O157: H7. Lancet 1987; ii: 276.
20.Levine, MM, Xu, JG. Kaper, JB et al. , A DNA probe to identify enterohemorrhagic Escherichia coli O157:H7 and other serotypes that cause hemorrhagic colitis and hemolytic uremic syndrome. J Infect Dis 1987; 156: 175–82.
21.Willshaw, GA, Smith, HR. Scotland, SM. Field, AM. Rowe, B. Heterogeneity of Escherichia coli phages encoding Vero cytotoxins: comparison of cloned sequences determining VT1 and VT2 and development of specific gene probes. J Gen Microbiol 1987; 133: 1309–17.
22.Moxley, RA, Francis, DH. Natural and experimental infection with an attaching and effacing strain of Escherichia coli in calves. Infect Immun 1986; 53: 339–46.
23.Tzipori, S. Wachsmuth, IK. Smithers, J. Jackson, C. Studies in gnotobiotic piglets on non-O157: H7 Escherichia coli serotypes isolated from patients with hemorrhagic colitis. Gastroenterol 1988; 94: 590–7.
24.Tzipori, S. Gibson, R. Montanaro, J. Nature and distribution of mucosal lesions associated with enteropathogenic and enterohemorrhagic Escherichia coli in piglets and the role of plasmid-mediated factors. Infect Immun 1989; 57: 1142–50.
25.Knutton, S. Baldwin, T. Williams, PH. McNeish, AS. Actin accumulation at sites of bacterial adhesion to tissue culture cells: basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli. Infect Immun 1989; 57: 1290–8.
26.Jerse, AE, Yu, J. Tall, BD, Kaper, JB. A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. Proc Natl Acad Sci USA 1990; 87: 7839–43.
27.Donnenberg, MS. Kaper, JB. Enteropathogenic Escherichia coli. Infect Immun 1992; 60: 3953–61.
28.Jerse, AE. Kaper, JB. The eae gene of enteropathogenie Escherichia coli encodes a 94-kilodalton membrane protein, the expression of which is influenced by the EAF plasmid. Infect Irnmun 1991; 59: 4302–9.
29.Beebakhee, G. Louie, M. De Azavedo, J. Brunton, J. Cloning and nucleotide sequence of the eae gene homologue from enterohemorrhagic Escherichia coli serotype O157: H7. FEMS Microbiol Lett 1992; 91: 63–8.
30.Yu, K. Kaper, JB. Cloning and characterization of the eae gene of enterohemorrhagic Escherichia coli O157: H7. Mol Microbiol 1992; 6: 411–17.
31.World Health Organization. Manual of investigation of acute enteric infections. Publication no. CDD/83.3 Rev. 1983: 1: 27.
32.Sambrook, J. Fritsch, EF. Maniatis, T. In: Molecular cloning. A laboratory manual. Nolan, C. ed. Cold Spring Harbor. NY: Cold Spring Harbor Laboratory Press, 1989.
33.Devereux, J. Haeberli, P. Smithies, O. A comprehensive set of sequence analysis programs for the VAN. Nucleic Acids Res 1984; 12: 387–95.
34.Read, SC. Clarke, RC. Martin, A. et al. Polymerase chain reaction for detection of veroeytotoxigenic Escherichia coli isolated from animal and food sources. Mol Cell Probes 1992; 6: 153–61.
35.Willshaw, GA. Scotland, SM. Smith, HR. Rowe, B. Properties of Yero cytotoxin-producing Escherichia coli of human origin of O serogroups other than O157: H7. J Infect Dis 1991; 166: 797802.
36.Merson, MH. Orskov, F. Orskov, I. Sack, R. Huq, I. Koster, F. Relationship between enterotoxin production and serotype in enterotoxigenic Escherichia coli. Infect Immun 1979; 23: 325–9.
37.Robins-Browne, RM. Traditional enteropathogenie Escherichia coli of infantile diarrhea. Rev Infect Dis 1987; 9: 2853.
38.Janke, BH. Francis, DH. Collins, JE. et al. Attaching and effacing Escherichia coli infection as a cause of diarrhea in young calves. J Am Yet Med Assoc 1990; 196: 897901.
39.Schoonderwoerd, M. Clarke, RC. van Dreumel, TA. Rawluk, SA. Colitis in calves: natural and experimental infection with a verotoxin-producing strain of Escherichia coli O111: NM. Can J Vet Res 1988; 52: 484–7.
40.Sherman, P. Soni, R. Karmali, M. Attaching and effacing adherence of verocytotoxin-producing Escherichia coli to rabbit intestinal epithelium in vivo. Infect Immun 1988; 56: 756–61.
41.Barrett, TJ. Kaper, JB. Jerse, AE. Wachsmuth, IK. Virulence factors in Shiga-like toxin-producing Escherichia coli isolated from humans and cattle. J Infect Dis 1992; 165: 979–80.
42.Griffin, PM. Tauxe, RV. The epidemiology of infections caused by Escherichia coli O157:H7. other enterohemorrhagic E. coli. and the associated hemolytic uremic syndrome. Epidemiol Rev 1991; 13: 6098.
43.Bertschinger, HU. Pohlenz, J. Bacterial colonization and morphology of the intestine in porcine Escherichia coli enterotoxemia (edema disease). Vet Pathol 1983; 20: 99100.
44.Gannon, VPJ. Rashed, M. King, RK. Golsteyn, Thomas EJ. Detection and characterization of the eae gene of Shiga-like toxin producing Escherichia coli using polymerase chain reaction. J Clin Microbiol 1993; 31: 1268–74.
45.Whittam, TS. Wolfe, ML. Wachsmuth, IK, Orskov, F. Orskov, I. Wilson, RA. Clonal relationships among Escherichia coli strains that cause hemorrhagic colitis and infantile diarrhea. Infect Immun 1993; 61: 1619–29.
46.Scotland, SM. Willshaw, GA. Smith, HR. Gross, RJ. Rowe, B. Adhesion to cells in culture and plasmid profiles of enteropathogenic Escherichia coli isolated from outbreaks and sporadic cases of infant diarrhoea. J Infect 1989; 19: 237–49.
47.Stenderup, J. Orskov, F. The clonal nature of enteropathogenie Escherichia coli strains. J Infect Dis 1983; 148: 1019–24.
48.Knutton, S. Shaw, RK. Bhan, MK. et al. Ability of enteroaggregative Escherichia coli strains to adhere in vitro to human intestinal mucosa. Infect Immun 1992; 60: 2086–91.
49.Scotland, SM. Smith, HR. Said, B. Willshaw, GA. Cheasty, T. Rowe, B. Identification of enteropathogenie Escherichia coli isolated in Britain as enteroaggregative or as members of a subclass of attaching-and-effacing E. coli not hybridising with the EPEC adherence-factor probe. J Med Microbiol 1991; 35: 278–83.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Epidemiology & Infection
  • ISSN: 0950-2688
  • EISSN: 1469-4409
  • URL: /core/journals/epidemiology-and-infection
Please enter your name
Please enter a valid email address
Who would you like to send this to? *


Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed