Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1987 Feb;25(2):278–284. doi: 10.1128/jcm.25.2.278-284.1987

Monoclonal antibody passive hemagglutination and capture enzyme-linked immunosorbent assays for direct detection and quantitation of F41 and K99 fimbrial antigens in enterotoxigenic Escherichia coli.

T J Raybould, C F Crouch, S D Acres
PMCID: PMC265883  PMID: 2880866

Abstract

Production of diarrhea in neonatal calves by enterotoxigenic Escherichia coli depends on its ability to attach to the epithelial cells of the intestine via surface adhesins called pili or fimbriae and to secrete enterotoxins. The most important of these fimbriae are designated K99 and F41. We produced and characterized a murine monoclonal antibody specific to F41. This monoclonal antibody and a K99-specific monoclonal antibody were used to develop sensitive and specific passive hemagglutination and capture enzyme-linked immunosorbent assays (ELISAs) for detection and quantitation of F41 and K99 antigens in E. coli cultures and culture supernatants. The capture ELISA systems exhibited excellent sensitivity and specificity, whereas the passive hemagglutination systems appeared to be oversensitive. The ability of the capture ELISAs to detect K99 and F41 fimbrial antigens in fecal specimens from calves was evaluated. Fimbrial antigens were detected in six of six specimens from scouring calves but not in four of four specimens from nonscouring calves.

Full text

PDF
278

Selected References

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

  1. Acres S. D., Isaacson R. E., Babiuk L. A., Kapitany R. A. Immunization of calves against enterotoxigenic colibacillosis by vaccinating dams with purified K99 antigen and whole cell bacterins. Infect Immun. 1979 Jul;25(1):121–126. doi: 10.1128/iai.25.1.121-126.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arbuckle J. B. The location of Escherichia coli in the pig intestine. J Med Microbiol. 1970 May;3(2):333–340. doi: 10.1099/00222615-3-2-333. [DOI] [PubMed] [Google Scholar]
  3. Bertschinger H. U., Moon H. W., Whipp S. C. Association of Escherichia coli with the small intestinal epithelium. I. Comparison of enteropathogenic and nonenteropathogenic porcine strains in pigs. Infect Immun. 1972 Apr;5(4):595–605. doi: 10.1128/iai.5.4.595-605.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burrows M. R., Sellwood R., Gibbons R. A. Haemagglutinating and adhesive properties associated with the K99 antigen of bovine strains of Escherichia coli. J Gen Microbiol. 1976 Oct;96(2):269–275. doi: 10.1099/00221287-96-2-269. [DOI] [PubMed] [Google Scholar]
  5. Chan R., Acres S. D., Costerton J. W. Use of specific antibody to demonstrate glycocalyx, K99 pili, and the spatial relationships of K99+ enterotoxigenic Escherichia coli in the ileum of colostrum-fed calves. Infect Immun. 1982 Sep;37(3):1170–1180. doi: 10.1128/iai.37.3.1170-1180.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Crouch C. F., Raybould T. J. Comparison of different antigen preparations as substrates for use in passive hemagglutination and enzyme-linked immunosorbent assays for detection of antibody against bovine enteric coronavirus. J Clin Microbiol. 1983 Jul;18(1):146–149. doi: 10.1128/jcm.18.1.146-149.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. DUGUID J. P., SMITH I. W., DEMPSTER G., EDMUNDS P. N. Non-flagellar filamentous appendages (fimbriae) and haemagglutinating activity in Bacterium coli. J Pathol Bacteriol. 1955 Oct;70(2):335–348. doi: 10.1002/path.1700700210. [DOI] [PubMed] [Google Scholar]
  8. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. J Immunol. 1972 Jul;109(1):129–135. [PubMed] [Google Scholar]
  9. Guinée P. A., Veldkamp J., Jansen W. H. Improved minca medium for the detection of K99 antigen in calf enterotoxigenic strains of Escherichia coli. Infect Immun. 1977 Feb;15(2):676–678. doi: 10.1128/iai.15.2.676-678.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Isaacson R. E. K99 surface antigen of Escherichia coli: purification and partial characterization. Infect Immun. 1977 Jan;15(1):272–279. doi: 10.1128/iai.15.1.272-279.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kennett R. H., Denis K. A., Tung A. S., Klinman N. R. Hybrid plasmacytoma production: fusions with adult spleen cells, monoclonal spleen fragments, neonatal spleen cells and human spleen cells. Curr Top Microbiol Immunol. 1978;81:77–91. doi: 10.1007/978-3-642-67448-8_13. [DOI] [PubMed] [Google Scholar]
  12. Mills K. W., Phillips R. M., Kelly B. L., Baughman G. L. Using enzyme-linked immunosorbent assay to detect Escherichia coli K88 pili antigens from clinical isolates. Am J Vet Res. 1982 Feb;43(2):365–367. [PubMed] [Google Scholar]
  13. Mills K. W., Tietze K. L. Monoclonal antibody enzyme-linked immunosorbent assay for identification of K99-positive Escherichia coli isolates from calves. J Clin Microbiol. 1984 Apr;19(4):498–501. doi: 10.1128/jcm.19.4.498-501.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mills K. W., Tietze K. L., Phillips R. M. Use of enzyme-linked immunosorbent assay for detection of K88 pili in fecal specimens from swine. Am J Vet Res. 1983 Nov;44(11):2188–2189. [PubMed] [Google Scholar]
  15. Mooi F. R., de Graaf F. K., van Embden J. D. Cloning, mapping and expression of the genetic determinant that encodes for the K88ab antigen. Nucleic Acids Res. 1979 Mar;6(3):849–865. doi: 10.1093/nar/6.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moon H. W. Mechanisms in the pathogenesis of diarrhea: a review. J Am Vet Med Assoc. 1978 Feb 15;172(4):443–448. [PubMed] [Google Scholar]
  17. Moon H. W., Nagy B., Isaacson R. E., Orskov I. Occurrence of K99 antigen on Escherichia coli isolated from pigs and colonization of pig ileum by K99+ enterotoxigenic E. coli from calves and pigs. Infect Immun. 1977 Feb;15(2):614–620. doi: 10.1128/iai.15.2.614-620.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moon H. W., Whipp S. C., Skartvedt S. M. Etiologic diagnosis of diarrheal disease of calves: frequency and methods for detecting enterotoxin and K99 antigen production by Escherichia cola. Am J Vet Res. 1976 Sep;37(9):1025–1029. [PubMed] [Google Scholar]
  19. Morris J. A., Thorns C. J., Sojka W. J. Evidence for two adhesive antigens on the K99 reference strain Escherichia coli B41. J Gen Microbiol. 1980 May;118(1):107–113. doi: 10.1099/00221287-118-1-107. [DOI] [PubMed] [Google Scholar]
  20. Morris J. A., Thorns C., Scott A. C., Sojka W. J., Wells G. A. Adhesion in vitro and in vivo associated with an adhesive antigen (F41) produced by a K99 mutant of the reference strain Escherichia coli B41. Infect Immun. 1982 Jun;36(3):1146–1153. doi: 10.1128/iai.36.3.1146-1153.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Myers L. L., Guinée P. A. Occurrence and characteristics of enterotoxigenic Escherichia coli isolated from calves with diarrhea. Infect Immun. 1976 Apr;13(4):1117–1119. doi: 10.1128/iai.13.4.1117-1119.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Myers L. L. Passive protection of calves against experimentally induced and naturally occurring enteric colibacillosis. Am J Vet Res. 1980 Dec;41(12):1952–1956. [PubMed] [Google Scholar]
  23. Nagy B. Vaccination of cows with a K99 extract to protect newborn calves against experimental enterotoxic colibacillosis. Infect Immun. 1980 Jan;27(1):21–24. doi: 10.1128/iai.27.1.21-24.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. O'Beirne A. J., Cooper H. R. Heterogeneous enzyme immunoassay. J Histochem Cytochem. 1979 Aug;27(8):1148–1162. doi: 10.1177/27.8.383822. [DOI] [PubMed] [Google Scholar]
  25. Orskov I., Orskov F., Smith H. W., Sojka W. J. The establishment of K99, a thermolabile, transmissible escherichia coli K antigen, previously called "Kco", possessed by calf and lamb enteropathogenic strains. Acta Pathol Microbiol Scand B. 1975 Feb;83(1):31–36. doi: 10.1111/j.1699-0463.1975.tb00066.x. [DOI] [PubMed] [Google Scholar]
  26. Raybould T. J., Chantler S. Serological differentiation between infected and vaccinated cattle by using purified soluble antigens from Brucella abortus in a hemagglutination system. Infect Immun. 1980 Aug;29(2):435–441. doi: 10.1128/iai.29.2.435-441.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Roberts P. L., Duncan B. E., Raybould T. J., Watson D. H. Purification of herpes simplex virus glycoproteins B and C using monoclonal antibodies and their ability to protect mice against lethal challenge. J Gen Virol. 1985 May;66(Pt 5):1073–1085. doi: 10.1099/0022-1317-66-5-1073. [DOI] [PubMed] [Google Scholar]
  28. Sequeira P. J., Eldridge A. E. Treponemal haemagglutination test. Br J Vener Dis. 1973 Jun;49(3):242–248. doi: 10.1136/sti.49.3.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sherman D. M., Acres S. D., Sadowski P. L., Springer J. A., Bray B., Raybould T. J., Muscoplat C. C. Protection of calves against fatal enteric colibacillosis by orally administered Escherichia coli K99-specific monoclonal antibody. Infect Immun. 1983 Nov;42(2):653–658. doi: 10.1128/iai.42.2.653-658.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Smith H. W., Halls S. Observations by the ligated intestinal segment and oral inoculation methods on Escherichia coli infections in pigs, calves, lambs and rabbits. J Pathol Bacteriol. 1967 Apr;93(2):499–529. doi: 10.1002/path.1700930211. [DOI] [PubMed] [Google Scholar]
  31. Snodgrass D. R., Nagy L. K., Sherwood D., Campbell I. Passive immunity in calf diarrhea: vaccination with K99 antigen of enterotoxigenic Escherichia coli and rotavirus. Infect Immun. 1982 Aug;37(2):586–591. doi: 10.1128/iai.37.2.586-591.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. To S. C. F41 antigen among porcine enterotoxigenic Escherichia coli strains lacking K88, K99, and 987P pili. Infect Immun. 1984 Feb;43(2):549–554. doi: 10.1128/iai.43.2.549-554.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Voller A., Bidwell D. E. A simple method for detecting antibodies to rubella. Br J Exp Pathol. 1975 Aug;56(4):338–339. [PMC free article] [PubMed] [Google Scholar]
  34. de Graaf F. K., Klemm P., Gaastra W. Purification, characterization, and partial covalent structure of Escherichia coli adhesive antigen K99. Infect Immun. 1981 Sep;33(3):877–883. doi: 10.1128/iai.33.3.877-883.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. de Graaf F. K., Roorda I. Production, purification, and characterization of the fimbrial adhesive antigen F41 isolated from calf enteropathogenic Escherichia coli strain B41M. Infect Immun. 1982 May;36(2):751–758. doi: 10.1128/iai.36.2.751-758.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES