Abstract
A total of 78 E. coli strains isolated from adults with different types of urinary tract infections were screened by polymerase chain reaction for prevalence of genetic regions coding for virulence factors. The targeted genetic determinants were those coding for type 1 fimbriae (fimH), pili associated with pyelonephritis (pap), S and F1C fimbriae (sfa and foc), afimbrial adhesins (afa), hemolysin (hly), cytotoxic necrotizing factor (cnf), aerobactin (aer). Among the studied strains, the prevalence of genes coding for fimbrial adhesive systems was 86 %, 36%, and 23% for fimH, pap, and sfa/foc, respectively. The operons coding for Afa afimbrial adhesins were identified in 14% of strains. The hly and cnf genes coding for toxins were amplified in 23% and 13% of strains, respectively. A prevalence of 54% was found for the aer gene. The various combinations of detected genes were designated as virulence patterns. The strains isolated from the hospitalized patients displayed a greater number of virulence genes and a diversity of gene associations compared to the strains isolated from the ambulatory subjects. A rapid assessment of the bacterial pathogenicity characteristics may contribute to a better medical approach of the patients with urinary tract infections.
Keywords: Escherichia coli, virulence factors, PCR, urinary tract infections
References
- 1. Donnenberg M.S., Welch R.A., Virulence determinants of uropathogenic Escherichia coli In: Urinary tract infections: molecular pathogenesis and clinical management (Mobley H., Warren J., eds.), 1996, pp. 135–174. ASM Press, Washington , D.C . [Google Scholar]
- 2. Zhang L., Foxman B., Manning S.D., Tallman P., Marrs C.F., Molecular epidemiologic approaches to urinary tract infection gene discovery in uropathogenic Escherichia coli, Infect. Immun., 68(4): 2009–2015, 2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3. Johnson J.R., Russo T.A., Tarr P.I., Carlino U., Bilge S.S., Vary J.C., Stell A.L., Molecular epidemiological and phylogenetic associations of two novel putative virulence genes, iha and iro NE.coli, among Escherichia coli isolates from patients with urosepsis, Infect. Immun., 68(5): 3040–3047, 2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4. Johnson J.R., Stell A.L., Extended virulence genotypes of Escherichia coli strains from patients with urosepsis in relation to phylogeny and host compromise, J. Infect. Dis., 181: 261–272, 2000. [DOI] [PubMed] [Google Scholar]
- 5. Foxman B., Zhang L., Tallman P., Palin K., Rode C., Bloch C., Gillespie B., Marrs C.F., Virulence characteristics of Escherichia coli causing first urinary tract infection predict risk of second infection, J. Infect. Dis., 172: 1536–1541, 1995. [DOI] [PubMed] [Google Scholar]
- 6. Johnson J.R., Brown J.J., Carlino U.B., Russo T.A., Colonization with and acquisition of uropathogenic Escherichia coli as revealed by polymerase chain reaction ‐ based detection, J. Infect. Dis., 177: 1120–1124, 1998. [DOI] [PubMed] [Google Scholar]
- 7. Le Bouguénec, C. , Archambaud M., Labigne A., Rapid and specific detection of the pap, afa, and sfa adhesins ‐ encoding operons in uropathogenic Escherichia coli strains by polymerase chain reaction, J. Clin. Microbiol., 30(5): 1189–1193, 1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8. Farmer J.J., Enterobacteriaceae: Introduction and identification In: Manual of clinical microbiology 7th ed.(Murray P.R., Baron E.J., Phaller M.A., Tenover F.C., Yolken R.H., eds), 1999, pp.442–458. ASM Press, Washington , D.C . [Google Scholar]
- 9. Le Bouguénec, C. , Lalioui L., Du Merle, L. , Jouve M., Courcoux P., Bouzari S., Selvarangan R., Nowicki B.J., Germani Y., Andremont A., Gounon P., Garcia M.I., Characterization of AfaE adhesins produced by extraintestinal and intestinal human Escherichia coli isolates. PCR assays for detection of Afa adhesins that do or do not recognize Dr blood group antigens, J. Clin. Microbiol., 39: 1738–1745, 2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10. Johnson J.R., Stell A.L., Extended virulence genotypes of Escherichia coli strains from patients with urosepsis in relation to phylogeny and host compromise, J. Infect. Dis., 181: 261–272, 2000. [DOI] [PubMed] [Google Scholar]
- 11. Bindereif A., Neilands J.B., Promotor mapping and transcriptional regulation of the iron assimilation system of plasmid ColV‐K30 in Escherichia coli K‐12, J. Bacteriol., 162: 1039–1046, 1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 12. Boyer H.W., Roulland‐Dussoix D., A complementation analysis of the restriction and modification of DNA in Escherichia coli, J. Mol. Biol., 41: 459–472, 1969. [DOI] [PubMed] [Google Scholar]
- 13. Blomfield I.C., McCain M.S., Einstein B.I., Type 1 fimbriae mutants of Escherichia coli K‐12: characterization of recognized afimbriate strains and construction of new deletion mutants, Mol. Microbiol., 5: 1439–1445, 1991. [DOI] [PubMed] [Google Scholar]
- 14. Stamm W.E., Hooton T.M., Management of urinary tract infections in adults, N. Engl. J. Med., 329(18): 1328–1334, 1993. [DOI] [PubMed] [Google Scholar]
- 15. Garcia M.‐I., Le Bouguénec, C. , Role of adhesion in pathogenicity of human uropathogenic and diarrhoeogenic Escherichia coli, Bull. Inst. Pasteur., 94: 201–236, 1996. [Google Scholar]
- 16. Connell H., Agace W., Klemm P., Schembri M., Marild S., Svanborg C., Type 1 fimbrial expression enhances Escherichia coli virulence for the urinary tract, Proc. Natl. Acad. Sci. USA, 93: 9827–9832, 1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 17. Martinez J.J., Mulvey M.A., Schilling J.D., Pinkner J.S., Hultgren S.J., Type 1 pilus‐mediated bacterial invasion of bladder epithelial cells, EMBO J., 19: 2803–2812, 2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 18. Johnson J.R., Virulence factors in Escherichia coli urinary tract infection, Clin. Microbiol. Rev., 4(1): 80–128, 1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 19. Sokurenko E.V., Chesnokova V., Dykhuizen D.E., Ofek I., Wu X.‐R., Krogfelt K.A., Struve C., Schembri M.A., Hasty D.L., Pathogenic adaptation of Escherichia coli by natural variation of the fimH adhesin, Proc. Natl. Acad. Sci. USA, 95: 8922–8926, 1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 20. Goluszko P., Moseley S., Truong L.D., Kaul A., Williford J.R., Selvarangan R., Nowicki S., Nowicki B., Development of experimental model of chronic pyelonephritis with Escherichia coli O75:K5:H‐bearing Dr fimbriae, J. Clin. Invest., 99: 1662–1672, 1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 21. Blanco J., Blanco M., Alonso M.P., Blanco J.E., Gonzalez E.A., Garabal J.I., Characteristics of haemolytic Escherichia coli with particular reference to production of cytotoxic necrotizing factor type 1 (CNF 1), Res. Microbiol., 143: 869–878, 1992. [DOI] [PubMed] [Google Scholar]
- 22. Hofman P., Flatau G., Selva E., Gauthier M., Le Negrate, G. , Fiorentini C., Rossi B., Boquet P., Escherichia coli cytotoxic necrotizing factor 1 effaces microvilli and decreases transmigration of polymorphonuclear leukocytes in intestinal T84 epithelial cell monolayers, Infect. Immun., 66(6): 2494–2500, 1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 23. Carbonetti N.H., Boonchai S., Parry S.H., Vaisanen‐Rhen V., Kornonen T.K., Williams P.H., Aerobactin‐mediated iron uptake by Escherichia coli isolates from human intestinal infections, Infect. Immun., 51: 966–968, 1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 24. Zhang L., Foxman B., Tallman P., Cladera E., Le Bouguénec, C. , Marrs C.F., Distribution of drb genes coding for Dr binding adhesins among uropathogenic and fecal Escherichia coli isolates and identification of new subtypes, Infect. Immun., 65(6): 2011–2018, 1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 25. Lalioui L., Le Bouguénec, C. , afa‐8 gene cluster is carried by a pathogenicity island inserted into the tRNA Phe of human and bovine pathogenic Escherichia coli isolates, Infect. Immun., 69: 937–948, 2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 26. Blum G., Falbo V., Caprioli A., Hacker J., Gene clusters encoding the cytotoxic necrotizing factor type 1, Prs fimbriae and ‐hemolysin form the pathogenicity island II of the uropathogenic Escherichia coli strain J96, FEMS Microbiol. Lett., 126: 189–196, 1995. [DOI] [PubMed] [Google Scholar]
- 27. Falbo V., Famiglietti M., Caprioli A., Gene block encoding production of cytotoxic necrotizing factor 1 and hemolysin in Escherichia coli isolates from extraintestinal infections, Infect. Immun., 60: 2182–2187, 1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 28. Hacker J., Bender L., Ott M., Wingender J., Lund B., Marre R., Goebel W., Deletions of chromosomal regions coding for fimbriae and hemolysins occur in vitro and in vivo in various extraintestinal Escherichia coli isolates, Microb. Pathog., 8: 213–225, 1990. [DOI] [PubMed] [Google Scholar]
- 29. Bingen E., Picard B., Brahimi N., Mathy S., Desjardins P., Elion J., Denamur E., Phylogenetic analysis of Escherichia coli strains causing neonatal meningitis suggests horizontal gene transfer from a predominant pool of highly virulent B2 group strains, J. Infect. Dis., 177: 642–650, 1998. [DOI] [PubMed] [Google Scholar]