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. 1996 Mar;34(3):689–694. doi: 10.1128/jcm.34.3.689-694.1996

Characteristics of Escherichia coli strains belonging to enteropathogenic E. coli serogroups isolated in Italy from children with diarrhea.

A Giammanco 1, M Maggio 1, G Giammanco 1, R Morelli 1, F Minelli 1, F Scheutz 1, A Caprioli 1
PMCID: PMC228871  PMID: 8904439

Abstract

Fifty-five Escherichia coli strains belonging to enteropathogenic E. coli (EPEC) serogroups were examined for phenotypic and genetic factors associated with virulence. The strains were isolated in Italy from children with diarrhea and identified as EPEC by clinical laboratories using commercially available antisera. O:H serotyping showed that 35 strains (27 of O26, O111, and O128 serogroups) belonged to 11 serotypes considered to be classical EPEC O:H serotypes. The other 20 isolates were classified as 15 nonclassical EPEC O:H serotypes. All the potential EPEC virulence factors associated with bacterial adhesion (localized adherence, fluorescentactin staining test positivity, presence of the attaching and effacing [eaeA] gene), the production of verotoxin, and the positivity with the enterohemorrhagic E. coli probe were significantly more frequent among isolates belonging to classical than nonclassical serotypes. Strains displaying an aggregative adhesion and hybridizing with the enteroaggregative DNA probe were found in serogroups O86, O111, and O126. Verotoxin-producing isolates belonged to serogroups O26, O111, and O128. Only one of the isolates hybridized with the EPEC adherence factor (EAF) probe, but 33 strains gave positive results with the eae probe, confirming that the former is more suitable in epidemiological studies in European countries. These results indicate that up to 75% of strains identified as EPEC by commercial antisera may possess potential virulence properties and/or belong to classical EPEC O:H serotypes and suggest that O grouping is still a useful diagnostic tool for presumptive identification of diarrheagenic E. coli in clinical laboratories.

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Selected References

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

  1. Albert M. J., Alam K., Ansaruzzaman M., Montanaro J., Islam M., Faruque S. M., Haider K., Bettelheim K., Tzipori S. Localized adherence and attaching-effacing properties of nonenteropathogenic serotypes of Escherichia coli. Infect Immun. 1991 May;59(5):1864–1868. doi: 10.1128/iai.59.5.1864-1868.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baudry B., Savarino S. J., Vial P., Kaper J. B., Levine M. M. A sensitive and specific DNA probe to identify enteroaggregative Escherichia coli, a recently discovered diarrheal pathogen. J Infect Dis. 1990 Jun;161(6):1249–1251. doi: 10.1093/infdis/161.6.1249. [DOI] [PubMed] [Google Scholar]
  3. Bhan M. K., Raj P., Levine M. M., Kaper J. B., Bhandari N., Srivastava R., Kumar R., Sazawal S. Enteroaggregative Escherichia coli associated with persistent diarrhea in a cohort of rural children in India. J Infect Dis. 1989 Jun;159(6):1061–1064. doi: 10.1093/infdis/159.6.1061. [DOI] [PubMed] [Google Scholar]
  4. Cantey J. R., Moseley S. L. HeLa cell adherence, actin aggregation, and invasion by nonenteropathogenic Escherichia coli possessing the eae gene. Infect Immun. 1991 Nov;59(11):3924–3929. doi: 10.1128/iai.59.11.3924-3929.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Caprioli A., Luzzi I., Rosmini F., Pasquini P., Cirrincione R., Gianviti A., Matteucci M. C., Rizzoni G. Hemolytic-uremic syndrome and Vero cytotoxin-producing Escherichia coli infection in Italy. The HUS Italian Study Group. J Infect Dis. 1992 Jul;166(1):154–158. doi: 10.1093/infdis/166.1.154. [DOI] [PubMed] [Google Scholar]
  6. Caprioli A., Luzzi I., Rosmini F., Resti C., Edefonti A., Perfumo F., Farina C., Goglio A., Gianviti A., Rizzoni G. Community-wide outbreak of hemolytic-uremic syndrome associated with non-O157 verocytotoxin-producing Escherichia coli. J Infect Dis. 1994 Jan;169(1):208–211. doi: 10.1093/infdis/169.1.208. [DOI] [PubMed] [Google Scholar]
  7. Donnenberg M. S., Kaper J. B. Enteropathogenic Escherichia coli. Infect Immun. 1992 Oct;60(10):3953–3961. doi: 10.1128/iai.60.10.3953-3961.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Donnenberg M. S., Tacket C. O., James S. P., Losonsky G., Nataro J. P., Wasserman S. S., Kaper J. B., Levine M. M. Role of the eaeA gene in experimental enteropathogenic Escherichia coli infection. J Clin Invest. 1993 Sep;92(3):1412–1417. doi: 10.1172/JCI116717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Donnenberg M. S., Yu J., Kaper J. B. A second chromosomal gene necessary for intimate attachment of enteropathogenic Escherichia coli to epithelial cells. J Bacteriol. 1993 Aug;175(15):4670–4680. doi: 10.1128/jb.175.15.4670-4680.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  11. Gannon V. P., Rashed M., King R. K., Thomas E. J. Detection and characterization of the eae gene of Shiga-like toxin-producing Escherichia coli using polymerase chain reaction. J Clin Microbiol. 1993 May;31(5):1268–1274. doi: 10.1128/jcm.31.5.1268-1274.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Girón J. A., Ho A. S., Schoolnik G. K. An inducible bundle-forming pilus of enteropathogenic Escherichia coli. Science. 1991 Nov 1;254(5032):710–713. doi: 10.1126/science.1683004. [DOI] [PubMed] [Google Scholar]
  13. Girón J. A., Ho A. S., Schoolnik G. K. Characterization of fimbriae produced by enteropathogenic Escherichia coli. J Bacteriol. 1993 Nov;175(22):7391–7403. doi: 10.1128/jb.175.22.7391-7403.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jerse A. E., Gicquelais K. G., Kaper J. B. Plasmid and chromosomal elements involved in the pathogenesis of attaching and effacing Escherichia coli. Infect Immun. 1991 Nov;59(11):3869–3875. doi: 10.1128/iai.59.11.3869-3875.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]
  18. 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]
  19. Knutton S., Phillips A. D., Smith H. R., Gross R. J., Shaw R., Watson P., Price E. Screening for enteropathogenic Escherichia coli in infants with diarrhea by the fluorescent-actin staining test. Infect Immun. 1991 Jan;59(1):365–371. doi: 10.1128/iai.59.1.365-371.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Knutton S., Shaw R. K., Bhan M. K., Smith H. R., McConnell M. M., Cheasty T., Williams P. H., Baldwin T. J. Ability of enteroaggregative Escherichia coli strains to adhere in vitro to human intestinal mucosa. Infect Immun. 1992 May;60(5):2083–2091. doi: 10.1128/iai.60.5.2083-2091.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Law D. Adhesion and its role in the virulence of enteropathogenic Escherichia coli. Clin Microbiol Rev. 1994 Apr;7(2):152–173. doi: 10.1128/cmr.7.2.152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Levine M. M., Nataro J. P., Karch H., Baldini M. M., Kaper J. B., Black R. E., Clements M. L., O'Brien A. D. The diarrheal response of humans to some classic serotypes of enteropathogenic Escherichia coli is dependent on a plasmid encoding an enteroadhesiveness factor. J Infect Dis. 1985 Sep;152(3):550–559. doi: 10.1093/infdis/152.3.550. [DOI] [PubMed] [Google Scholar]
  23. Levine M. M., Prado V., Robins-Browne R., Lior H., Kaper J. B., Moseley S. L., Gicquelais K., Nataro J. P., Vial P., Tall B. Use of DNA probes and HEp-2 cell adherence assay to detect diarrheagenic Escherichia coli. J Infect Dis. 1988 Jul;158(1):224–228. doi: 10.1093/infdis/158.1.224. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Louie M., de Azavedo J., Clarke R., Borczyk A., Lior H., Richter M., Brunton J. Sequence heterogeneity of the eae gene and detection of verotoxin-producing Escherichia coli using serotype-specific primers. Epidemiol Infect. 1994 Jun;112(3):449–461. doi: 10.1017/s0950268800051153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Morelli R., Baldassarri L., Falbo V., Donelli G., Caprioli A. Detection of enteroadherent Escherichia coli associated with diarrhoea in Italy. J Med Microbiol. 1994 Dec;41(6):399–404. doi: 10.1099/00222615-41-6-399. [DOI] [PubMed] [Google Scholar]
  27. Morris K. J., Rao G. G. Conventional screening for enteropathogenic Escherichia coli in the UK. Is it appropriate or necessary? J Hosp Infect. 1992 Jul;21(3):163–167. doi: 10.1016/0195-6701(92)90072-t. [DOI] [PubMed] [Google Scholar]
  28. Nataro J. P., Baldini M. M., Kaper J. B., Black R. E., Bravo N., Levine M. M. Detection of an adherence factor of enteropathogenic Escherichia coli with a DNA probe. J Infect Dis. 1985 Sep;152(3):560–565. doi: 10.1093/infdis/152.3.560. [DOI] [PubMed] [Google Scholar]
  29. Nataro J. P., Kaper J. B., Robins-Browne R., Prado V., Vial P., Levine M. M. Patterns of adherence of diarrheagenic Escherichia coli to HEp-2 cells. Pediatr Infect Dis J. 1987 Sep;6(9):829–831. doi: 10.1097/00006454-198709000-00008. [DOI] [PubMed] [Google Scholar]
  30. Pedroso M. Z., Freymüller E., Trabulsi L. R., Gomes T. A. Attaching-effacing lesions and intracellular penetration in HeLa cells and human duodenal mucosa by two Escherichia coli strains not belonging to the classical enteropathogenic E. coli serogroups. Infect Immun. 1993 Mar;61(3):1152–1156. doi: 10.1128/iai.61.3.1152-1156.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Robins-Browne R. M., Yam W. C., O'Gorman L. E., Bettelheim K. A. Examination of archetypal strains of enteropathogenic Escherichia coli for properties associated with bacterial virulence. J Med Microbiol. 1993 Mar;38(3):222–226. doi: 10.1099/00222615-38-3-222. [DOI] [PubMed] [Google Scholar]
  32. Savarino S. J., Fasano A., Watson J., Martin B. M., Levine M. M., Guandalini S., Guerry P. Enteroaggregative Escherichia coli heat-stable enterotoxin 1 represents another subfamily of E. coli heat-stable toxin. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):3093–3097. doi: 10.1073/pnas.90.7.3093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schmidt H., Rüssmann H., Karch H. Virulence determinants in nontoxinogenic Escherichia coli O157 strains that cause infantile diarrhea. Infect Immun. 1993 Nov;61(11):4894–4898. doi: 10.1128/iai.61.11.4894-4898.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Scotland S. M., Smith H. R., Rowe B. Escherichia coli O128 strains from infants with diarrhea commonly show localized adhesion and positivity in the fluorescent-actin staining test but do not hybridize with an enteropathogenic E. coli adherence factor probe. Infect Immun. 1991 Apr;59(4):1569–1571. doi: 10.1128/iai.59.4.1569-1571.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Scotland S. M., Smith H. R., Said B., Willshaw G. A., Cheasty T., Rowe B. Identification of enteropathogenic 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 Nov;35(5):278–283. doi: 10.1099/00222615-35-5-278. [DOI] [PubMed] [Google Scholar]
  36. Scotland S. M., Willshaw G. A., Smith H. R., Rowe B. Properties of strains of Escherichia coli O26:H11 in relation to their enteropathogenic or enterohemorrhagic classification. J Infect Dis. 1990 Nov;162(5):1069–1074. doi: 10.1093/infdis/162.5.1069. [DOI] [PubMed] [Google Scholar]
  37. Smith H. R., Scotland S. M., Stokes N., Rowe B. Examination of strains belonging to enteropathogenic Escherichia coli serogroups for genes encoding EPEC adherence factor and Vero cytotoxins. J Med Microbiol. 1990 Apr;31(4):235–240. doi: 10.1099/00222615-31-4-235. [DOI] [PubMed] [Google Scholar]
  38. Smith H. R., Scotland S. M., Willshaw G. A., Rowe B., Cravioto A., Eslava C. Isolates of Escherichia coli O44:H18 of diverse origin are enteroaggregative. J Infect Dis. 1994 Dec;170(6):1610–1613. doi: 10.1093/infdis/170.6.1610. [DOI] [PubMed] [Google Scholar]
  39. Tesh V. L., O'Brien A. D. Adherence and colonization mechanisms of enteropathogenic and enterohemorrhagic Escherichia coli. Microb Pathog. 1992 Apr;12(4):245–254. doi: 10.1016/0882-4010(92)90043-n. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. Willshaw G. A., Scotland S. M., Smith H. R., Rowe B. Properties of Vero cytotoxin-producing Escherichia coli of human origin of O serogroups other than O157. J Infect Dis. 1992 Oct;166(4):797–802. doi: 10.1093/infdis/166.4.797. [DOI] [PubMed] [Google Scholar]
  42. Willshaw G. A., Smith H. R., Scotland S. M., Field A. M., 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 May;133(5):1309–1317. doi: 10.1099/00221287-133-5-1309. [DOI] [PubMed] [Google Scholar]
  43. 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]

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