Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1993 Mar;31(3):558–564. doi: 10.1128/jcm.31.3.558-564.1993

Characterization of monoclonal antibodies against putative colonization factors of enterotoxigenic Escherichia coli and their use in an epidemiological study.

G I Viboud 1, N Binsztein 1, A M Svennerholm 1
PMCID: PMC262819  PMID: 8096215

Abstract

Monoclonal antibodies (MAbs) against five putative colonization factors (PCFs), i.e., colonization factor antigen (CFA)/III, coli surface antigen (CS)7 and CS17, PCFO159, and PCFO166 of enterotoxigenic Escherichia coli (ETEC), were produced. Hybridomas (one each) producing specific antibodies against the respective PCFs were selected. All the MAbs reacted with the corresponding fimbriae but not with CFA/I, CFA/II, or CFA/IV or the heterologous PCFs in bacterial agglutination and enzyme-linked immunosorbent assays (ELISAs). In immunoelectron microscopy these MAbs bound along the fimbriae, and they also reacted with the corresponding subunits in immunoblots. The five MAbs were used to evaluate the prevalence of CFA/III, CS7, CS17, PCFO159, and PCFO166 in ETEC strains isolated from children with diarrhea in Argentina. One hundred five ETEC isolates negative for CFA/I, CFA/II, and CFA/IV were tested in slide agglutination or in a dot blot test for spontaneously agglutinating strains; positive results were confirmed by inhibition ELISAs. It was found that 27% of the CFA-negative ETEC strains carried one of the PCFs. The sensitivity of slide agglutination with these MAbs was similar to that with specific polyclonal antisera; however, the specificity was higher. PCFO166 was found in 9.5% of the strains tested, mainly in ETEC of serogroup O78 producing heat-stable toxin alone. CS17 and CS7 were identified in 6.7 and 5.7%, respectively, of strains producing heat-labile toxin only, most of which belonged to serogroup O114. PCFO159 was found in 3.8% of the isolates tested, whereas CFA/III was detected in only one ETEC strain.

Full text

PDF

Images in this article

560Image
on p.560
561Image
on p.561
561Image
on p.561
562Image
on p.562

Selected References

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

  1. Agüero M. E., Reyes L., Prado V., Orskov I., Orskov F., Cabello F. C. Enterotoxigenic Escherichia coli in a population of infants with diarrhea in Chile. J Clin Microbiol. 1985 Oct;22(4):576–581. doi: 10.1128/jcm.22.4.576-581.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aubel D., Darfeuille-Michaud A., Joly B. New adhesive factor (antigen 8786) on a human enterotoxigenic Escherichia coli O117:H4 strain isolated in Africa. Infect Immun. 1991 Apr;59(4):1290–1299. doi: 10.1128/iai.59.4.1290-1299.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Binsztein N., Jouve M. J., Viboud G. I., López Moral L., Rivas M., Orskov I., Ahrén C., Svennerholm A. M. Colonization factors of enterotoxigenic Escherichia coli isolated from children with diarrhea in Argentina. J Clin Microbiol. 1991 Sep;29(9):1893–1898. doi: 10.1128/jcm.29.9.1893-1898.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Changchawalit S., Echeverria P., Taylor D. N., Leksomboon U., Tirapat C., Eampokalap B., Rowe B. Colonization factors associated with enterotoxigenic Escherichia coli isolated in Thailand. Infect Immun. 1984 Aug;45(2):525–527. doi: 10.1128/iai.45.2.525-527.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Darfeuille-Michaud A., Forestier C., Joly B., Cluzel R. Identification of a nonfimbrial adhesive factor of an enterotoxigenic Escherichia coli strain. Infect Immun. 1986 May;52(2):468–475. doi: 10.1128/iai.52.2.468-475.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Deneke C. F., Thorne G. M., Gorbach S. L. Serotypes of attachment pili of enterotoxigenic Escherichia coli isolated from humans. Infect Immun. 1981 Jun;32(3):1254–1260. doi: 10.1128/iai.32.3.1254-1260.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Evans D. G., Evans D. J., Jr, Clegg S., Pauley J. A. Purification and characterization of the CFA/I antigen of enterotoxigenic Escherichia coli. Infect Immun. 1979 Aug;25(2):738–748. doi: 10.1128/iai.25.2.738-748.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Evans D. G., Evans D. J., Jr New surface-associated heat-labile colonization factor antigen (CFA/II) produced by enterotoxigenic Escherichia coli of serogroups O6 and O8. Infect Immun. 1978 Aug;21(2):638–647. doi: 10.1128/iai.21.2.638-647.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Evans D. G., Silver R. P., Evans D. J., Jr, Chase D. G., Gorbach S. L. Plasmid-controlled colonization factor associated with virulence in Esherichia coli enterotoxigenic for humans. Infect Immun. 1975 Sep;12(3):656–667. doi: 10.1128/iai.12.3.656-667.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gothefors L., Ahrén C., Stoll B., Barua D. K., Orskov F., Salek M. A., Svennerholm A. M. Presence of colonization factor antigens on fresh isolates of fecal Escherichia coli: a prospective study. J Infect Dis. 1985 Dec;152(6):1128–1133. doi: 10.1093/infdis/152.6.1128. [DOI] [PubMed] [Google Scholar]
  11. Heuzenroeder M. W., Elliot T. R., Thomas C. J., Halter R., Manning P. A. A new fimbrial type (PCFO9) on enterotoxigenic Escherichia coli 09:H- LT+ isolated from a case of infant diarrhea in central Australia. FEMS Microbiol Lett. 1990 Jan 1;54(1-3):55–60. doi: 10.1016/0378-1097(90)90258-r. [DOI] [PubMed] [Google Scholar]
  12. Hibberd M. L., McConnell M. M., Field A. M., Rowe B. The fimbriae of human enterotoxigenic Escherichia coli strain 334 are related to CS5 fimbriae. J Gen Microbiol. 1990 Dec;136(12):2449–2456. doi: 10.1099/00221287-136-12-2449. [DOI] [PubMed] [Google Scholar]
  13. Honda T., Arita M., Miwatani T. Characterization of new hydrophobic pili of human enterotoxigenic Escherichia coli: a possible new colonization factor. Infect Immun. 1984 Mar;43(3):959–965. doi: 10.1128/iai.43.3.959-965.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Knutton S., Lloyd D. R., Candy D. C., McNeish A. S. Ultrastructural study of adhesion of enterotoxigenic Escherichia coli to erythrocytes and human intestinal epithelial cells. Infect Immun. 1984 May;44(2):519–527. doi: 10.1128/iai.44.2.519-527.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Knutton S., Lloyd D. R., McNeish A. S. Identification of a new fimbrial structure in enterotoxigenic Escherichia coli (ETEC) serotype O148:H28 which adheres to human intestinal mucosa: a potentially new human ETEC colonization factor. Infect Immun. 1987 Jan;55(1):86–92. doi: 10.1128/iai.55.1.86-92.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lopez-Vidal Y., Klemm P., Svennerholm A. M. Monoclonal antibodies against different epitopes on colonization factor antigen I of enterotoxin-producing Escherichia coli. J Clin Microbiol. 1988 Oct;26(10):1967–1972. doi: 10.1128/jcm.26.10.1967-1972.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. López-Vidal Y., Calva J. J., Trujillo A., Ponce de León A., Ramos A., Svennerholm A. M., Ruiz-Palacios G. M. Enterotoxins and adhesins of enterotoxigenic Escherichia coli: are they risk factors for acute diarrhea in the community? J Infect Dis. 1990 Aug;162(2):442–447. doi: 10.1093/infdis/162.2.442. [DOI] [PubMed] [Google Scholar]
  19. Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
  20. McConnell M. M., Chart H., Field A. M., Hibberd M., Rowe B. Characterization of a putative colonization factor (PCFO166) of enterotoxigenic Escherichia coli of serogroup O166. J Gen Microbiol. 1989 May;135(5):1135–1144. doi: 10.1099/00221287-135-5-1135. [DOI] [PubMed] [Google Scholar]
  21. McConnell M. M., Hibberd M. L., Penny M. E., Scotland S. M., Cheasty T., Rowe B. Surveys of human enterotoxigenic Escherichia coli from three different geographical areas for possible colonization factors. Epidemiol Infect. 1991 Jun;106(3):477–484. doi: 10.1017/s0950268800067522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McConnell M. M., Hibberd M., Field A. M., Chart H., Rowe B. Characterization of a new putative colonization factor (CS17) from a human enterotoxigenic Escherichia coli of serotype O114:H21 which produces only heat-labile enterotoxin. J Infect Dis. 1990 Feb;161(2):343–347. doi: 10.1093/infdis/161.2.343. [DOI] [PubMed] [Google Scholar]
  23. McConnell M. M., Rowe B. Prevalence of the putative colonization factors CFA/III and PCFO159:H4 in enterotoxigenic Escherichia coli. J Infect Dis. 1989 Mar;159(3):582–586. doi: 10.1093/infdis/159.3.582. [DOI] [PubMed] [Google Scholar]
  24. McConnell M. M., Thomas L. V., Day N. P., Rowe B. Enzyme-linked immunosorbent assays for the detection of adhesion factor antigens of enterotoxigenic Escherichia coli. J Infect Dis. 1985 Dec;152(6):1120–1127. doi: 10.1093/infdis/152.6.1120. [DOI] [PubMed] [Google Scholar]
  25. McConnell M. M., Thomas L. V., Willshaw G. A., Smith H. R., Rowe B. Genetic control and properties of coli surface antigens of colonization factor antigen IV (PCF8775) of enterotoxigenic Escherichia coli. Infect Immun. 1988 Aug;56(8):1974–1980. doi: 10.1128/iai.56.8.1974-1980.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Smyth C. J. Two mannose-resistant haemagglutinins on enterotoxigenic Escherichia coli of serotype O6:K15:H16 or H-isolated from travellers' and infantile diarrhoea. J Gen Microbiol. 1982 Sep;128(9):2081–2096. doi: 10.1099/00221287-128-9-2081. [DOI] [PubMed] [Google Scholar]
  27. Svennerholm A. M., Holmgren J., Sack D. A. Development of oral vaccines against enterotoxinogenic Escherichia coli diarrhoea. Vaccine. 1989 Jun;7(3):196–198. doi: 10.1016/0264-410x(89)90228-4. [DOI] [PubMed] [Google Scholar]
  28. Tacket C. O., Maneval D. R., Levine M. M. Purification, morphology, and genetics of a new fimbrial putative colonization factor of enterotoxigenic Escherichia coli O159:H4. Infect Immun. 1987 May;55(5):1063–1069. doi: 10.1128/iai.55.5.1063-1069.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Thomas L. V., McConnell M. M., Rowe B., Field A. M. The possession of three novel coli surface antigens by enterotoxigenic Escherichia coli strains positive for the putative colonization factor PCF8775. J Gen Microbiol. 1985 Sep;131(9):2319–2326. doi: 10.1099/00221287-131-9-2319. [DOI] [PubMed] [Google Scholar]

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

RESOURCES