Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1993 May;31(5):1268–1274. doi: 10.1128/jcm.31.5.1268-1274.1993

Detection and characterization of the eae gene of Shiga-like toxin-producing Escherichia coli using polymerase chain reaction.

V P Gannon 1, M Rashed 1, R K King 1, E J Thomas 1
PMCID: PMC262916  PMID: 8501228

Abstract

In this study, the polymerase chain reaction (PCR) was used in the detection of the attaching and effacing (eae) gene of Shiga-like toxin-producing Escherichia coli (SLT-EC). Oligonucleotide primers, complementary to the 5' portion of the eae gene of the enteropathogenic E. coli E2348/69 (O127:H6) and of SLT-EC CL8 and EDL933 (O157:H7), generated PCR products of the predicted sizes with DNA from the majority of human clinical SLT-EC strains tested from O serogroups 5, 26, 103, 111, 121, 128, 145, and 157; all SLT-EC strains of O serogroups 5, 26, and 111 from cattle; and a minority of porcine SLT-EC strains (one strain each from O serogroups 107 and 130 and one rough strain). Five HaeIII digestion profiles were obtained for PCR products generated by amplification of a 2.3-kb DNA fragment from the 5' end of eae. The HaeIII profiles for SLT-EC O serogroups, such as 26, 103, and 157, differed from each other but were consistent among strains within these O serogroups. Oligonucleotide primer pairs complementary to the 3' end of either the O127:H6 E. coli or the O157:H7 eae nucleotide sequence only amplified DNA from E. coli strains from a few of the SLT-EC O serogroups examined. One primer pair with homology to the 3' nucleotide sequence of eae from E. coli O157:H7 appeared to be relatively specific for this O serogroup by PCR. No PCR products were obtained in amplification experiments with the eae primers using DNA from human SLT-EC of O serogroups 38 (1 0f 1) and 91 (3 or 3), 15 of 15 SLT-EC strains from edema disease, or 29 of 29 non-SLT-EC strains from pigs and calves with diarrhea.

Full text

PDF
1268

Images in this article

1271Image
on p.1271
1272Image
on p.1272
1272Image
on p.1272

Selected References

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

  1. Barrett T. J., Kaper J. B., Jerse A. E., Wachsmuth I. K. Virulence factors in Shiga-like toxin-producing Escherichia coli isolated from humans and cattle. J Infect Dis. 1992 May;165(5):979–980. doi: 10.1093/infdis/165.5.979. [DOI] [PubMed] [Google Scholar]
  2. 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 Feb 1;70(1):63–68. doi: 10.1016/0378-1097(92)90563-4. [DOI] [PubMed] [Google Scholar]
  3. Borczyk A. A., Karmali M. A., Lior H., Duncan L. M. Bovine reservoir for verotoxin-producing Escherichia coli O157:H7. Lancet. 1987 Jan 10;1(8524):98–98. doi: 10.1016/s0140-6736(87)91928-3. [DOI] [PubMed] [Google Scholar]
  4. Chanter N., Hall G. A., Bland A. P., Hayle A. J., Parsons K. R. Dysentery in calves caused by an atypical strain of Escherichia coli (S102-9). Vet Microbiol. 1986 Sep;12(3):241–253. doi: 10.1016/0378-1135(86)90053-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chart H., Scotland S. M., Willshaw G. A., Rowe B. HEp-2 adhesion and the expression of a 94 kDa outer-membrane protein by strains of Escherichia coli belonging to enteropathogenic serogroups. J Gen Microbiol. 1988 May;134(5):1315–1321. doi: 10.1099/00221287-134-5-1315. [DOI] [PubMed] [Google Scholar]
  6. Clarke R. C., McEwen S. A., Gannon V. P., Lior H., Gyles C. L. Isolation of verocytotoxin-producing Escherichia coli from milk filters in south-western Ontario. Epidemiol Infect. 1989 Apr;102(2):253–260. doi: 10.1017/s0950268800029927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Doyle M. P., Schoeni J. L. Isolation of Escherichia coli O157:H7 from retail fresh meats and poultry. Appl Environ Microbiol. 1987 Oct;53(10):2394–2396. doi: 10.1128/aem.53.10.2394-2396.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gannon V. P., Gyles C. L. Characteristics of the Shiga-like toxin produced by Escherichia coli associated with porcine edema disease. Vet Microbiol. 1990 Jul;24(1):89–100. doi: 10.1016/0378-1135(90)90054-y. [DOI] [PubMed] [Google Scholar]
  9. Jerse A. E., Kaper J. B. The eae gene of enteropathogenic Escherichia coli encodes a 94-kilodalton membrane protein, the expression of which is influenced by the EAF plasmid. Infect Immun. 1991 Dec;59(12):4302–4309. doi: 10.1128/iai.59.12.4302-4309.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jerse A. E., Yu J., Tall B. D., Kaper J. B. A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. Proc Natl Acad Sci U S A. 1990 Oct;87(20):7839–7843. doi: 10.1073/pnas.87.20.7839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Karmali M. A. Infection by verocytotoxin-producing Escherichia coli. Clin Microbiol Rev. 1989 Jan;2(1):15–38. doi: 10.1128/cmr.2.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Karmali M. A., Petric M., Lim C., Fleming P. C., Arbus G. S., Lior H. The association between idiopathic hemolytic uremic syndrome and infection by verotoxin-producing Escherichia coli. J Infect Dis. 1985 May;151(5):775–782. doi: 10.1093/infdis/151.5.775. [DOI] [PubMed] [Google Scholar]
  13. Knutton S., Baldwin T., Williams P. H., McNeish A. S. 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 Apr;57(4):1290–1298. doi: 10.1128/iai.57.4.1290-1298.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Knutton S., Lloyd D. R., McNeish A. S. Adhesion of enteropathogenic Escherichia coli to human intestinal enterocytes and cultured human intestinal mucosa. Infect Immun. 1987 Jan;55(1):69–77. doi: 10.1128/iai.55.1.69-77.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Konowalchuk J., Speirs J. I., Stavric S. Vero response to a cytotoxin of Escherichia coli. Infect Immun. 1977 Dec;18(3):775–779. doi: 10.1128/iai.18.3.775-779.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Levine M. M., Xu J. G., Kaper J. B., Lior H., Prado V., Tall B., Nataro J., Karch H., Wachsmuth K. A DNA probe to identify enterohemorrhagic Escherichia coli of O157:H7 and other serotypes that cause hemorrhagic colitis and hemolytic uremic syndrome. J Infect Dis. 1987 Jul;156(1):175–182. doi: 10.1093/infdis/156.1.175. [DOI] [PubMed] [Google Scholar]
  17. Mainil J. G., Duchesnes C. J., Whipp S. C., Marques L. R., O'Brien A. D., Casey T. A., Moon H. W. Shiga-like toxin production and attaching effacing activity of Escherichia coli associated with calf diarrhea. Am J Vet Res. 1987 May;48(5):743–748. [PubMed] [Google Scholar]
  18. Martin M. L., Shipman L. D., Wells J. G., Potter M. E., Hedberg K., Wachsmuth I. K., Tauxe R. V., Davis J. P., Arnoldi J., Tilleli J. Isolation of Escherichia coli O157:H7 from dairy cattle associated with two cases of haemolytic uraemic syndrome. Lancet. 1986 Nov 1;2(8514):1043–1043. doi: 10.1016/s0140-6736(86)92656-5. [DOI] [PubMed] [Google Scholar]
  19. Moon H. W., Whipp S. C., Argenzio R. A., Levine M. M., Giannella R. A. Attaching and effacing activities of rabbit and human enteropathogenic Escherichia coli in pig and rabbit intestines. Infect Immun. 1983 Sep;41(3):1340–1351. doi: 10.1128/iai.41.3.1340-1351.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. O'Brien A. D., Holmes R. K. Shiga and Shiga-like toxins. Microbiol Rev. 1987 Jun;51(2):206–220. doi: 10.1128/mr.51.2.206-220.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. O'Brien A. D., LaVeck G. D., Thompson M. R., Formal S. B. Production of Shigella dysenteriae type 1-like cytotoxin by Escherichia coli. J Infect Dis. 1982 Dec;146(6):763–769. doi: 10.1093/infdis/146.6.763. [DOI] [PubMed] [Google Scholar]
  22. Orskov F., Orskov I., Villar J. A. Cattle as reservoir of verotoxin-producing Escherichia coli O157:H7. Lancet. 1987 Aug 1;2(8553):276–276. doi: 10.1016/s0140-6736(87)90860-9. [DOI] [PubMed] [Google Scholar]
  23. Read S. C., Gyles C. L., Clarke R. C., Lior H., McEwen S. Prevalence of verocytotoxigenic Escherichia coli in ground beef, pork, and chicken in southwestern Ontario. Epidemiol Infect. 1990 Aug;105(1):11–20. doi: 10.1017/s0950268800047592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Riley L. W., Remis R. S., Helgerson S. D., McGee H. B., Wells J. G., Davis B. R., Hebert R. J., Olcott E. S., Johnson L. M., Hargrett N. T. Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med. 1983 Mar 24;308(12):681–685. doi: 10.1056/NEJM198303243081203. [DOI] [PubMed] [Google Scholar]
  25. Schoonderwoerd M., Clarke R. C., van Dreumel A. A., Rawluk S. A. Colitis in calves: natural and experimental infection with a verotoxin-producing strain of Escherichia coli O111:NM. Can J Vet Res. 1988 Oct;52(4):484–487. [PMC free article] [PubMed] [Google Scholar]
  26. Sherman P., Cockerill F., 3rd, Soni R., Brunton J. Outer membranes are competitive inhibitors of Escherichia coli O157:H7 adherence to epithelial cells. Infect Immun. 1991 Mar;59(3):890–899. doi: 10.1128/iai.59.3.890-899.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sherman P., Soni R., Karmali M. Attaching and effacing adherence of Vero cytotoxin-producing Escherichia coli to rabbit intestinal epithelium in vivo. Infect Immun. 1988 Apr;56(4):756–761. doi: 10.1128/iai.56.4.756-761.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sherwood D., Snodgrass D. R., O'Brien A. D. Shiga-like toxin production from Escherichia coli associated with calf diarrhoea. Vet Rec. 1985 Feb 23;116(8):217–218. doi: 10.1136/vr.116.8.217. [DOI] [PubMed] [Google Scholar]
  29. Smith H. W., Green P., Parsell Z. Vero cell toxins in Escherichia coli and related bacteria: transfer by phage and conjugation and toxic action in laboratory animals, chickens and pigs. J Gen Microbiol. 1983 Oct;129(10):3121–3137. doi: 10.1099/00221287-129-10-3121. [DOI] [PubMed] [Google Scholar]
  30. Thomas E. J., King R. K., Burchak J., Gannon V. P. Sensitive and specific detection of Listeria monocytogenes in milk and ground beef with the polymerase chain reaction. Appl Environ Microbiol. 1991 Sep;57(9):2576–2580. doi: 10.1128/aem.57.9.2576-2580.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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 Apr;57(4):1142–1150. doi: 10.1128/iai.57.4.1142-1150.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tzipori S., Wachsmuth I. K., Chapman C., Birden R., Brittingham J., Jackson C., Hogg J. The pathogenesis of hemorrhagic colitis caused by Escherichia coli O157:H7 in gnotobiotic piglets. J Infect Dis. 1986 Oct;154(4):712–716. doi: 10.1093/infdis/154.4.712. [DOI] [PubMed] [Google Scholar]
  33. Wells J. G., Davis B. R., Wachsmuth I. K., Riley L. W., Remis R. S., Sokolow R., Morris G. K. Laboratory investigation of hemorrhagic colitis outbreaks associated with a rare Escherichia coli serotype. J Clin Microbiol. 1983 Sep;18(3):512–520. doi: 10.1128/jcm.18.3.512-520.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wells J. G., Shipman L. D., Greene K. D., Sowers E. G., Green J. H., Cameron D. N., Downes F. P., Martin M. L., Griffin P. M., Ostroff S. M. Isolation of Escherichia coli serotype O157:H7 and other Shiga-like-toxin-producing E. coli from dairy cattle. J Clin Microbiol. 1991 May;29(5):985–989. doi: 10.1128/jcm.29.5.985-989.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yu J., Kaper J. B. Cloning and characterization of the eae gene of enterohaemorrhagic Escherichia coli O157:H7. Mol Microbiol. 1992 Feb;6(3):411–417. doi: 10.1111/j.1365-2958.1992.tb01484.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES