Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1991 Aug;59(8):2664–2672. doi: 10.1128/iai.59.8.2664-2672.1991

Virulence patterns and long-range genetic mapping of extraintestinal Escherichia coli K1, K5, and K100 isolates: use of pulsed-field gel electrophoresis.

M Ott 1, L Bender 1, G Blum 1, M Schmittroth 1, M Achtman 1, H Tschäpe 1, J Hacker 1
PMCID: PMC258071  PMID: 1677349

Abstract

A total of 127 extraintestinal Escherichia coli strains of the capsule serotypes K1, K5, and K100 from human and animal sources were analyzed for DNA sequences specific for the genes for various adhesins (P fimbriae [pap] and P-related sequences [prs], S fimbriae [sfa]/F1C fimbriae [foc], and type I fimbriae [fim]), aerobactin (aer), and hemolysin (hly). The expression of corresponding virulence factors was also tested. Twenty-four selected strains were analyzed by long-range DNA mapping to evaluate their genetic relationships. DNA sequences for the adhesins were often found in strains not expressing them, while strains with hemolysin and aerobactin genes usually did express them. Different isolates of the same serotype often expressed different virulence patterns. The use of virulence-associated gene probes for Southern hybridization with genomic DNA fragments separated by pulsed-field gel electrophoresis revealed that a highly heterogeneous restriction fragment length and hybridization pattern existed even within strains of the same serotype. Long-range DNA mapping is therefore useful for the evaluation of genetic relatedness among individual isolates and facilitates the performance of precise molecular epidemiology.

Full text

PDF
2665

Images in this article

2668Image
on p.2668
2669Image
on p.2669

Selected References

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

  1. Abraham J. M., Freitag C. S., Clements J. R., Eisenstein B. I. An invertible element of DNA controls phase variation of type 1 fimbriae of Escherichia coli. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5724–5727. doi: 10.1073/pnas.82.17.5724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Achtman M., Heuzenroeder M., Kusecek B., Ochman H., Caugant D., Selander R. K., Väisanen-Rhen V., Korhonen T. K., Stuart S., Orskov F. Clonal analysis of Escherichia coli O2:K1 isolated from diseased humans and animals. Infect Immun. 1986 Jan;51(1):268–276. doi: 10.1128/iai.51.1.268-276.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Achtman M., Mercer A., Kusecek B., Pohl A., Heuzenroeder M., Aaronson W., Sutton A., Silver R. P. Six widespread bacterial clones among Escherichia coli K1 isolates. Infect Immun. 1983 Jan;39(1):315–335. doi: 10.1128/iai.39.1.315-335.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Achtman M., Pluschke G. Clonal analysis of descent and virulence among selected Escherichia coli. Annu Rev Microbiol. 1986;40:185–210. doi: 10.1146/annurev.mi.40.100186.001153. [DOI] [PubMed] [Google Scholar]
  5. Arbeit R. D., Arthur M., Dunn R., Kim C., Selander R. K., Goldstein R. Resolution of recent evolutionary divergence among Escherichia coli from related lineages: the application of pulsed field electrophoresis to molecular epidemiology. J Infect Dis. 1990 Feb;161(2):230–235. doi: 10.1093/infdis/161.2.230. [DOI] [PubMed] [Google Scholar]
  6. Arthur M., Johnson C. E., Rubin R. H., Arbeit R. D., Campanelli C., Kim C., Steinbach S., Agarwal M., Wilkinson R., Goldstein R. Molecular epidemiology of adhesin and hemolysin virulence factors among uropathogenic Escherichia coli. Infect Immun. 1989 Feb;57(2):303–313. doi: 10.1128/iai.57.2.303-313.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bender L., Ott M., Marre R., Hacker J. Genome analysis of Legionella ssp. by orthogonal field alternation gel electrophoresis (OFAGE). FEMS Microbiol Lett. 1990 Nov;60(3):253–257. doi: 10.1016/0378-1097(90)90313-f. [DOI] [PubMed] [Google Scholar]
  8. Braun V., Gross R., Köster W., Zimmermann L. Plasmid and chromosomal mutants in the iron(III)-aerobactin transport system of Escherichia coli. Use of streptonigrin for selection. Mol Gen Genet. 1983;192(1-2):131–139. doi: 10.1007/BF00327658. [DOI] [PubMed] [Google Scholar]
  9. Carbonetti N. H., Boonchai S., Parry S. H., Väisänen-Rhen V., Korhonen T. K., Williams P. H. Aerobactin-mediated iron uptake by Escherichia coli isolates from human extraintestinal infections. Infect Immun. 1986 Mar;51(3):966–968. doi: 10.1128/iai.51.3.966-968.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cross A. S., Kim K. S., Wright D. C., Sadoff J. C., Gemski P. Role of lipopolysaccharide and capsule in the serum resistance of bacteremic strains of Escherichia coli. J Infect Dis. 1986 Sep;154(3):497–503. doi: 10.1093/infdis/154.3.497. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Gross R., Engelbrecht F., Braun V. Identification of the genes and their polypeptide products responsible for aerobactin synthesis by pColV plasmids. Mol Gen Genet. 1985;201(2):204–212. doi: 10.1007/BF00425661. [DOI] [PubMed] [Google Scholar]
  13. Göransson M., Forsman K., Uhlin B. E. Functional and structural homology among regulatory cistrons of pili-adhesin determinants in Escherichia coli. Mol Gen Genet. 1988 Jun;212(3):412–417. doi: 10.1007/BF00330844. [DOI] [PubMed] [Google Scholar]
  14. 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. 1990 Mar;8(3):213–225. doi: 10.1016/0882-4010(90)90048-u. [DOI] [PubMed] [Google Scholar]
  15. Hacker J. Genetic determinants coding for fimbriae and adhesins of extraintestinal Escherichia coli. Curr Top Microbiol Immunol. 1990;151:1–27. doi: 10.1007/978-3-642-74703-8_1. [DOI] [PubMed] [Google Scholar]
  16. Hacker J., Hughes C. Genetics of Escherichia coli hemolysin. Curr Top Microbiol Immunol. 1985;118:139–162. doi: 10.1007/978-3-642-70586-1_8. [DOI] [PubMed] [Google Scholar]
  17. Hacker J., Schmidt G., Hughes C., Knapp S., Marget M., Goebel W. Cloning and characterization of genes involved in production of mannose-resistant, neuraminidase-susceptible (X) fimbriae from a uropathogenic O6:K15:H31 Escherichia coli strain. Infect Immun. 1985 Feb;47(2):434–440. doi: 10.1128/iai.47.2.434-440.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hacker J., Schröter G., Schrettenbrunner A., Hughes C., Goebel W. Hemolytic Escherichia coli strains in the human fecal flora as potential urinary pathogens. Zentralbl Bakteriol Mikrobiol Hyg A. 1983 May;254(3):370–378. [PubMed] [Google Scholar]
  19. High N. J., Hales B. A., Jann K., Boulnois G. J. A block of urovirulence genes encoding multiple fimbriae and hemolysin in Escherichia coli O4:K12:H-. Infect Immun. 1988 Feb;56(2):513–517. doi: 10.1128/iai.56.2.513-517.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hull S. I., Bieler S., Hull R. A. Restriction fragment length polymorphism and multiple copies of DNA sequences homologous with probes for P-fimbriae and hemolysin genes among uropathogenic Escherichia coli. Can J Microbiol. 1988 Mar;34(3):307–311. doi: 10.1139/m88-056. [DOI] [PubMed] [Google Scholar]
  21. Johnson J. R., Moseley S. L., Roberts P. L., Stamm W. E. Aerobactin and other virulence factor genes among strains of Escherichia coli causing urosepsis: association with patient characteristics. Infect Immun. 1988 Feb;56(2):405–412. doi: 10.1128/iai.56.2.405-412.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Klemm P. Two regulatory fim genes, fimB and fimE, control the phase variation of type 1 fimbriae in Escherichia coli. EMBO J. 1986 Jun;5(6):1389–1393. doi: 10.1002/j.1460-2075.1986.tb04372.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Knapp S., Hacker J., Then I., Müller D., Goebel W. Multiple copies of hemolysin genes and associated sequences in the chromosomes of uropathogenic Escherichia coli strains. J Bacteriol. 1984 Sep;159(3):1027–1033. doi: 10.1128/jb.159.3.1027-1033.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Korhonen T. K., Valtonen M. V., Parkkinen J., Väisänen-Rhen V., Finne J., Orskov F., Orskov I., Svenson S. B., Mäkelä P. H. Serotypes, hemolysin production, and receptor recognition of Escherichia coli strains associated with neonatal sepsis and meningitis. Infect Immun. 1985 May;48(2):486–491. doi: 10.1128/iai.48.2.486-491.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Korhonen T. K., Väisänen-Rhen V., Rhen M., Pere A., Parkkinen J., Finne J. Escherichia coli fimbriae recognizing sialyl galactosides. J Bacteriol. 1984 Aug;159(2):762–766. doi: 10.1128/jb.159.2.762-766.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kusecek B., Wloch H., Mercer A., Vaisänen V., Pluschke G., Korhonen T., Achtman M. Lipopolysaccharide, capsule, and fimbriae as virulence factors among O1, O7, O16, O18, or O75 and K1, K5, or K100 Escherichia coli. Infect Immun. 1984 Jan;43(1):368–379. doi: 10.1128/iai.43.1.368-379.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lafont J. P., Dho M., D'Hauteville H. M., Bree A., Sansonetti P. J. Presence and expression of aerobactin genes in virulent avian strains of Escherichia coli. Infect Immun. 1987 Jan;55(1):193–197. doi: 10.1128/iai.55.1.193-197.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lai E., Birren B. W., Clark S. M., Simon M. I., Hood L. Pulsed field gel electrophoresis. Biotechniques. 1989 Jan;7(1):34–42. [PubMed] [Google Scholar]
  29. Linggood M. A., Roberts M., Ford S., Parry S. H., Williams P. H. Incidence of the aerobactin iron uptake system among Escherichia coli isolates from infections of farm animals. J Gen Microbiol. 1987 Apr;133(4):835–842. doi: 10.1099/00221287-133-4-835. [DOI] [PubMed] [Google Scholar]
  30. Low D. A., Braaten B. A., Ling G. V., Johnson D. L., Ruby A. L. Isolation and comparison of Escherichia coli strains from canine and human patients with urinary tract infections. Infect Immun. 1988 Oct;56(10):2601–2609. doi: 10.1128/iai.56.10.2601-2609.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Low D., David V., Lark D., Schoolnik G., Falkow S. Gene clusters governing the production of hemolysin and mannose-resistant hemagglutination are closely linked in Escherichia coli serotype O4 and O6 isolates from urinary tract infections. Infect Immun. 1984 Jan;43(1):353–358. doi: 10.1128/iai.43.1.353-358.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Marre R., Hacker J. Role of S- and common-type I-fimbriae of Escherichia coli in experimental upper and lower urinary tract infection. Microb Pathog. 1987 Mar;2(3):223–226. doi: 10.1016/0882-4010(87)90023-4. [DOI] [PubMed] [Google Scholar]
  33. Mercer A. A., Morelli G., Heuzenroeder M., Kamke M., Achtman M. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins. Infect Immun. 1984 Dec;46(3):649–657. doi: 10.1128/iai.46.3.649-657.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Noegel A., Rdest U., Goebel W. Determination of the functions of hemolytic plasmid pHly152 of Escherichia coli. J Bacteriol. 1981 Jan;145(1):233–247. doi: 10.1128/jb.145.1.233-247.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Normark S., Lark D., Hull R., Norgren M., Båga M., O'Hanley P., Schoolnik G., Falkow S. Genetics of digalactoside-binding adhesin from a uropathogenic Escherichia coli strain. Infect Immun. 1983 Sep;41(3):942–949. doi: 10.1128/iai.41.3.942-949.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nowicki B., Vuopio-Varkila J., Viljanen P., Korhonen T. K., Mäkelä P. H. Fimbrial phase variation and systemic E. coli infection studied in the mouse peritonitis model. Microb Pathog. 1986 Aug;1(4):335–347. doi: 10.1016/0882-4010(86)90066-5. [DOI] [PubMed] [Google Scholar]
  37. Orskov I., Orskov F. Serology of Escherichia coli fimbriae. Prog Allergy. 1983;33:80–105. [PubMed] [Google Scholar]
  38. Orskov I., Williams P. H., Svanborg Edén C., Orskov F. Assessment of biological and colony hybridization assays for detection of the aerobactin system in Escherichia coli from urinary tract infections. Med Microbiol Immunol. 1989;178(3):143–148. doi: 10.1007/BF00198013. [DOI] [PubMed] [Google Scholar]
  39. Ott M., Hacker J., Schmoll T., Jarchau T., Korhonen T. K., Goebel W. Analysis of the genetic determinants coding for the S-fimbrial adhesin (sfa) in different Escherichia coli strains causing meningitis or urinary tract infections. Infect Immun. 1986 Dec;54(3):646–653. doi: 10.1128/iai.54.3.646-653.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ott M., Hoschützky H., Jann K., Van Die I., Hacker J. Gene clusters for S fimbrial adhesin (sfa) and F1C fimbriae (foc) of Escherichia coli: comparative aspects of structure and function. J Bacteriol. 1988 Sep;170(9):3983–3990. doi: 10.1128/jb.170.9.3983-3990.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Ott M., Schmoll T., Goebel W., Van Die I., Hacker J. Comparison of the genetic determinant coding for the S-fimbrial adhesin (sfa) of Escherichia coli to other chromosomally encoded fimbrial determinants. Infect Immun. 1987 Aug;55(8):1940–1943. doi: 10.1128/iai.55.8.1940-1943.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Parkkinen J., Finne J., Achtman M., Väisänen V., Korhonen T. K. Escherichia coli strains binding neuraminyl alpha 2-3 galactosides. Biochem Biophys Res Commun. 1983 Mar 16;111(2):456–461. doi: 10.1016/0006-291x(83)90328-5. [DOI] [PubMed] [Google Scholar]
  43. Parkkinen J., Rogers G. N., Korhonen T., Dahr W., Finne J. Identification of the O-linked sialyloligosaccharides of glycophorin A as the erythrocyte receptors for S-fimbriated Escherichia coli. Infect Immun. 1986 Oct;54(1):37–42. doi: 10.1128/iai.54.1.37-42.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Pawelzik M., Heesemann J., Hacker J., Opferkuch W. Cloning and characterization of a new type of fimbria (S/F1C-related fimbria) expressed by an Escherichia coli O75:K1:H7 blood culture isolate. Infect Immun. 1988 Nov;56(11):2918–2924. doi: 10.1128/iai.56.11.2918-2924.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Pere A., Leinonen M., Väisänen-Rhen V., Rhen M., Korhonen T. K. Occurrence of type-1C fimbriae on Escherichia coli strains isolated from human extraintestinal infections. J Gen Microbiol. 1985 Jul;131(7):1705–1711. doi: 10.1099/00221287-131-7-1705. [DOI] [PubMed] [Google Scholar]
  46. Robbins J. B., McCracken G. H., Jr, Gotschlich E. C., Orskov F., Orskov I., Hanson L. A. Escherichia coli K1 capsular polysaccharide associated with neonatal meningitis. N Engl J Med. 1974 May 30;290(22):1216–1220. doi: 10.1056/NEJM197405302902202. [DOI] [PubMed] [Google Scholar]
  47. Roberts M., Roberts I., Korhonen T. K., Jann K., Bitter-Suermann D., Boulnois G. J., Williams P. H. DNA probes for K-antigen (capsule) typing of Escherichia coli. J Clin Microbiol. 1988 Feb;26(2):385–387. doi: 10.1128/jcm.26.2.385-387.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Schmoll T., Morschhäuser J., Ott M., Ludwig B., van Die I., Hacker J. Complete genetic organization and functional aspects of the Escherichia coli S fimbrial adhesion determinant: nucleotide sequence of the genes sfa B, C, D, E, F. Microb Pathog. 1990 Nov;9(5):331–343. doi: 10.1016/0882-4010(90)90067-z. [DOI] [PubMed] [Google Scholar]
  49. Selander R. K., Korhonen T. K., Väisänen-Rhen V., Williams P. H., Pattison P. E., Caugant D. A. Genetic relationships and clonal structure of strains of Escherichia coli causing neonatal septicemia and meningitis. Infect Immun. 1986 Apr;52(1):213–222. doi: 10.1128/iai.52.1.213-222.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  51. Svanborg Edén C., de Man P. Bacterial virulence in urinary tract infection. Infect Dis Clin North Am. 1987 Dec;1(4):731–750. [PubMed] [Google Scholar]
  52. Valvano M. A., Silver R. P., Crosa J. H. Occurrence of chromosome- or plasmid-mediated aerobactin iron transport systems and hemolysin production among clonal groups of human invasive strains of Escherichia coli K1. Infect Immun. 1986 Apr;52(1):192–199. doi: 10.1128/iai.52.1.192-199.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Virkola R., Westerlund B., Holthöfer H., Parkkinen J., Kekomäki M., Korhonen T. K. Binding characteristics of Escherichia coli adhesins in human urinary bladder. Infect Immun. 1988 Oct;56(10):2615–2622. doi: 10.1128/iai.56.10.2615-2622.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Väisänen-Rhen V., Elo J., Väisänen E., Siitonen A., Orskov I., Orskov F., Svenson S. B., Mäkelä P. H., Korhonen T. K. P-fimbriated clones among uropathogenic Escherichia coli strains. Infect Immun. 1984 Jan;43(1):149–155. doi: 10.1128/iai.43.1.149-155.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Whittam T. S., Wilson R. A. Genetic relationships among pathogenic strains of avian Escherichia coli. Infect Immun. 1988 Sep;56(9):2458–2466. doi: 10.1128/iai.56.9.2458-2466.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wittig W., Prager R., Tietze E., Seltmann G., Tschäpe H. Aerobactin-positive Escherichia coli as causative agents of extra-intestinal infections among animals. Arch Exp Veterinarmed. 1988 Mar;42(2):221–229. [PubMed] [Google Scholar]
  57. van Die I., van Geffen B., Hoekstra W., Bergmans H. Type 1C fimbriae of a uropathogenic Escherichia coli strain: cloning and characterization of the genes involved in the expression of the 1C antigen and nucleotide sequence of the subunit gene. Gene. 1985;34(2-3):187–196. doi: 10.1016/0378-1119(85)90127-1. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES