1887

Abstract

The aim of this study was to investigate the role and clinical course of verotoxigenic (VTEC) infections in children with acute diarrhoea from Argentina, the country with the highest worldwide incidence of haemolytic uraemic syndrome (HUS). To accomplish this objective, 437 samples from children up to 6 years old with acute diarrhoea were collected and processed. More than 60 % of the children studied presented watery or mucous diarrhoea without blood, and in 25.2 % of the cases the samples contained blood. In a first screening, a multiplex PCR was performed to detect the presence of the , , , and virulence genes. The strains were then isolated and analysed to characterize their serotypes, virulence genes, antibiotic susceptibility profiles and verotoxin (VT) production. Forty-four of the 437 samples (10.1 %) were positive for VTEC virulence genes. VTEC-infected patients presented different types of diarrhoea (27.3 % belonged to the non-bloody type). Several serotypes and virulence genotypes were found. Isolates belonged to the serotypes O157 : H7, O145 : H, O26 : H11, O121 : H19, O111 : H2 and O118 : H2. HUS developed in 16 (36.4 %) patients positive for VTEC virulence genes. All of the VTEC isolates produced a cytopathic effect on Vero cell monolayers, confirming the ability to express VT. Despite most strains being sensitive to all of the antimicrobials studied, a positive association between clinical progression to HUS and antibiotic therapy was observed for the total number of patients studied, as well as for the VTEC group. In conclusion, the data obtained in this study increase our knowledge of the role and clinical course of VTEC infection in childhood acute diarrhoea beyond bloody diarrhoea, and might be considered for the prevention, diagnosis and management of this disease. It is possible that the optimal approach for VTEC diagnosis could be using multiplex PCR to search for the presence of the , , and genes.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.015560-0
2010-03-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jmm/59/3/345.html?itemId=/content/journal/jmm/10.1099/jmm.0.015560-0&mimeType=html&fmt=ahah

References

  1. Banatvala N., Griffin P. M., Greene K. D., Barrett T. J., Bibb W. F., Green J. H., Wells J. G. Hemolytic Uremic Syndrome Study Collaborators 2001; The United States National Prospective Hemolytic Uremic Syndrome Study: microbiologic, serologic, clinical, and epidemiologic findings. J Infect Dis 183:1063–1070 [CrossRef]
    [Google Scholar]
  2. Bauer A. W., Kirby W. M. M., Sherris J. C., Turck M. 1966; Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 45:493–496
    [Google Scholar]
  3. Beutin L., Zimmermann S., Gleier K. 1998; Human infections with Shiga toxin-producing Escherichia coli other than serogroup O157 in Germany. Emerg Infect Dis 4:635–639 [CrossRef]
    [Google Scholar]
  4. Blanco J., Blanco M., Alonso M. P., Blanco J. E., Garabal J. I., González E. A. 1992; Serogroups of Escherichia coli strains producing cytotoxic necrotizing factors CNF1 and CNF2. FEMS Microbiol Lett 75:155–159
    [Google Scholar]
  5. Blanco J. E., Blanco M., Mora A., Prado C., Río M., Fernández L., Fernández M. J., Saínz V., Blanco J. 1996; Detection of enterohaemorrhagic Escherichia coli O157 : H7 in ground beef using immunomagnetic separation. Microbiologia 12:385–394
    [Google Scholar]
  6. Blanco M., Blanco J. E., Blanco J., Mora A., Prado C., Alonso M. P., Mouriño M., Madrid C., Balsalobre C. other authors 1997; Distribution and characterization of fecal verotoxin-producing Escherichia coli (VTEC) isolated from healthy cattle. Comparison with VTEC obtained from calves with diarrhea. Vet Microbiol 54:309–319 [CrossRef]
    [Google Scholar]
  7. Blanco J. E., Blanco M., Alonso M. P., Mora A., Dahbi G., Coira M. A., Blanco J. 2004; Serotypes, virulence genes, and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolates from human patients: prevalence in Lugo, Spain, from 1992 through 1999. J Clin Microbiol 42:311–319 [CrossRef]
    [Google Scholar]
  8. Chen J., Griffiths M. W. 1998; PCR differentiation of Escherichia coli from other Gram-negative bacteria using primers derived from the nucleotide sequences flanking the gene encoding the universal stress protein. Lett Appl Microbiol 27:369–371 [CrossRef]
    [Google Scholar]
  9. CLSI 2008; Performance Standards for Antimicrobial Susceptibility Testing , 18th Informational Supplement. M100-S18 Wayne, PA: Clinical and Laboratory Standards Institute;
  10. Conover W. J. 1971 Practical Nonparametric Statistics New York: Wiley;
    [Google Scholar]
  11. Elliott E. J., Robins-Browne R. M., O’Loughlin E. V., Bennett-Wood V., Bourke J., Henning P., Hogg G. G., Knight J., Powell H. other authors 2001; Nationwide study of haemolytic uraemic syndrome: clinical, microbiological, and epidemiological features. Arch Dis Child 85:125–131 [CrossRef]
    [Google Scholar]
  12. Espié E., Grimont F., Mariani-Kurkdjian P., Bouvet P., Haeghebaert S., Filliol I., Loirat C., Decludt B., Nhu Tran Minh N. other authors 2008; Surveillance of hemolytic uremic syndrome in children less than 15 years of age, a system to monitor O157 and non-O157 Shiga toxin-producing Escherichia coli infections in France, 1996–2000. Pediatr Infect Dis J 27:595–601 [CrossRef]
    [Google Scholar]
  13. Gannon V. P. J., Rashed M., King R. K., Golsteyn Thomas E. J. 1993; Detection and characterization of the eae gene of Shiga-like toxin-producing Escherichia coli using polymerase chain reaction. J Clin Microbiol 31:1268–1274
    [Google Scholar]
  14. Gerber A., Karch H., Allerberg F., Verweyen M., Zimmerhackl L. B. 2002; Clinical course and the role of Shiga toxin-producing Escherichia coli infection in the hemolytic-uremic syndrome in pediatric patients, 1997–2000, in Germany and Austria: a prospective study. J Infect Dis 186:493–500 [CrossRef]
    [Google Scholar]
  15. Giugno S. M., Bibiloni N., Rahman R., Miliwebsky E., Chinen I., Rivas M. 2007; Association between uremic hemolytic syndrome and infection by Shiga toxin-producing Escherichia coli . Acta Bioquím Clín Latinoam 41:27–33 (in Spanish
    [Google Scholar]
  16. Goldwater P. N. 2007; Treatment and prevention of enterohemorrhagic Escherichia coli infection and hemolytic uremic syndrome. Expert Rev Anti Infect Ther 5:653–663 [CrossRef]
    [Google Scholar]
  17. Goldwater P. N., Bettelheim K. A. 1998; New perspectives on the role of Escherichia coli O157 : H7 and other enterohaemorrhagic E. coli serotypes in human disease. J Med Microbiol 47:1039–1045 [CrossRef]
    [Google Scholar]
  18. Griffin P. M., Tauxe R. V. 1991; The epidemiology of infections caused by Escherichia coli O157 : H7, other enterohemorrhagic E. coli , and the associated hemolytic uremic syndrome. Epidemiol Rev 13:60–98
    [Google Scholar]
  19. Johnson K. E., Thorpe C. M., Sears C. L. 2006; The emerging clinical importance of non-O157 Shiga toxin-producing Escherichia coli . Clin Infect Dis 43:1587–1595 [CrossRef]
    [Google Scholar]
  20. Karch H., Tarr P. I., Bielaszewska M. 2005; Enterohaemorrhagic Escherichia coli in human medicine. Int J Med Microbiol 295:405–418 [CrossRef]
    [Google Scholar]
  21. Karmali M. A. 1989; Infection by verocytotoxin-producing Escherichia coli . Clin Microbiol Rev 2:15–38
    [Google Scholar]
  22. Klein E. J., Stapp J. R., Clausen C. R., Boster D. R., Wells J. G., Qin X., Swerdlow D. L., Tarr P. I. 2002; Shiga toxin-producing Escherichia coli in children with diarrhea: a prospective point-of-care study. J Pediatr 141:172–177 [CrossRef]
    [Google Scholar]
  23. Krüger A., Padola N. L., Parma A. E., Lucchesi P. M. A. 2006; Intraserotype diversity among Argentinian verocytotoxigenic Escherichia coli detected by random amplified polymorphic DNA analysis. J Med Microbiol 55:545–549 [CrossRef]
    [Google Scholar]
  24. Lindqvist R., Antonsson A. K., Norling B., Persson L., Ekström C. L., Fäger U., Eriksson E., Lofdahl S., Norberg P. 1998; The prevalence of verocytotoxin-producing Escherichia coli (VTEC) and E. coli O157 : H7 in beef in Sweden determined by PCR assays and an immuno-magnetic separation (IMS) method. Food Microbiol 15:591–601 [CrossRef]
    [Google Scholar]
  25. López E. L., Contrini M. M., De Rosa M. F. 1998; Epidemiology of Shiga toxin-producing E. coli in South America. In Escherichia coli O157  :  H7 and Other Shiga Toxin-Producing E. coli Strains pp 30–37 Edited by Kaper J. B., O’Brien A. D. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  26. Orskov F., Orskov I. 1984; Serotyping of Escherichia coli . Methods Microbiol 114:43–112
    [Google Scholar]
  27. Oshima T. 1997; Predictive factors for development of hemolytic uremic syndrome (HUS) and early intensive treatments for prevention of HUS enterohemorrhagic Escherichia coli infection. Jpn J Antibiot 50:855–861
    [Google Scholar]
  28. Padola N. L., Sanz M. E., Blanco J. E., Blanco M., Blanco J., Etcheverría A. I., Arroyo G. H., Usera M. A., Parma A. E. 2004; Serotypes and virulence genes of Shigatoxigenic Escherichia coli (STEC) isolated from a feedlot in Argentina. Vet Microbiol 100:3–9 [CrossRef]
    [Google Scholar]
  29. Paton A. W., Paton J. C. 1998; Detection and characterization of Shiga toxigenic Escherichia coli by using multiplex PCR assays forstx 1, stx 2, eae ,enterohemorrhagic E. coli hlyA , rfb O111,and rfb O157. J Clin Microbiol 36:598–602
    [Google Scholar]
  30. Paton A. W., Paton J. C. 2002; Direct detection and characterization of Shiga toxigenic Escherichia coli by multiplex PCR for stx 1,stx 2, eae , ehxA ,and saa . J Clin Microbiol 40:271–274 [CrossRef]
    [Google Scholar]
  31. Paton A. W., Ratcliff R. M., Doyle R. M., Seymour-Murray J., Davos D., Lanser J. A., Paton J. C. 1996; Molecular microbiological investigation of an outbreak of hemolytic-uremic syndrome caused by dry fermented sausage contaminated with Shiga-like toxin-producing Escherichia coli . J Clin Microbiol 34:1622–1627
    [Google Scholar]
  32. Paton A. W., Srimanote P., Woodrow M. C., Paton J. C. 2001; Characterization of Saa, a novel autoagglutinating adhesin produced by locus of enterocyte effacement-negative Shiga-toxigenic Escherichia coli strains that are virulent for humans. Infect Immun 69:6999–7009 [CrossRef]
    [Google Scholar]
  33. Piérard D., Stevens D., Moriau L., Lior H., Lauwers S. 1997; Isolation and virulence factors of verocytotoxin-producing Escherichia coli in human stool samples. Clin Microbiol Infect 3:531–540 [CrossRef]
    [Google Scholar]
  34. Pradel N., Livrelli V., De Champs C., Palcoux J.-B., Reynaud A., Scheutz F., Sirot J., Joly B., Forestier C. 2000; Prevalence and characterization of Shiga toxin-producing Escherichia coli isolated from cattle, food and children during a one-year prospective study in France. J Clin Microbiol 38:1023–1031
    [Google Scholar]
  35. Proulx F., Seidman E. G., Karpman D. K. 2001; Pathogenesis of Shiga toxin-associated hemolytic uremic syndrome. Pediatr Res 50:163–171 [CrossRef]
    [Google Scholar]
  36. Rivas M., Miliwebsky E., Chinen I., Deza N., Leotta G. A. 2006; The epidemiology of hemolytic uremic syndrome in Argentina. Diagnosis of the etiologic agent, reservoirs and routes of transmission. Medicina (B Aires) 66 (Suppl. 3):27–32 (in Spanish
    [Google Scholar]
  37. Rivas M., Sosa-Estani S., Rangel J., Caletti M. G., Vallés P., Roldán C. D., Balbi L., Marsano de Mollar M. C., Amoedo D. other authors 2008; Risk factors for sporadic Shiga toxin-producing Escherichia coli infections in children, Argentina. Emerg Infect Dis 14:763–771 [CrossRef]
    [Google Scholar]
  38. Rivero M. A., Padola N. L., Etcheverría A. I., Parma A. E. 2004; Enterohemorrhagic Escherichia coli and hemolytic-uremic syndrome in Argentina. Medicina (B Aires) 64:352–356 (in Spanish
    [Google Scholar]
  39. Sanz M. E., Viñas M. R., Parma A. E. 1998; Prevalence of bovine verotoxin-producing Escherichia coli in Argentina. Eur J Epidemiol 14:399–403 [CrossRef]
    [Google Scholar]
  40. Slutsker L., Ries A. A., Greene K. D., Wells J. G., Hutwagner L., Griffin P. 1997; Escherichia coli O157 : H7 diarrhea in the United States: clinical and epidemiological features. Ann Intern Med 126:505–513 [CrossRef]
    [Google Scholar]
  41. Tarr P. I., Gordon C. A., Chandler W. L. 2005; Shiga-toxin-producing Escherichia coli and haemolytic uraemic syndrome. Lancet 365:1073–1086
    [Google Scholar]
  42. Voyer L. E. 1996 Síndrome Urémico Hemolítico Buenos Aires: Ed. López;
    [Google Scholar]
  43. Willshaw G. A., Cheasty T., Smith H. R., O’Brien S. J., Adak G. K. 2001; Verocytotoxin-producing Escherichia coli (VTEC) O157 and other VTEC from human infections in England and Wales: 1995–1998. J Med Microbiol 50:135–142
    [Google Scholar]
  44. Wong C. S., Jelacic S., Habeeb R. L., Watkins S. L., Tarr P. I. 2000; The risk of the hemolytic-uremic syndrome after antibiotic treatment of Escherichia coli O157 : H7 infections. N Engl J Med 342:1930–1936 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.015560-0
Loading
/content/journal/jmm/10.1099/jmm.0.015560-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error