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. 1986 Mar;23(3):586–591. doi: 10.1128/jcm.23.3.586-591.1986

Comparison of methods for detection of colonization factor antigens on enterotoxigenic Escherichia coli.

C M Ahrén, L Gothefors, B J Stoll, M A Salek, A M Svennerholm
PMCID: PMC268699  PMID: 3082927

Abstract

Fecal Escherichia coli isolates from 196 patients with watery diarrhea and 68 healthy individuals (controls) were analyzed in Bangladesh immediately after isolation for the presence of colonization factor antigen (CFA) I or II (CFA/I or CFA/II, respectively) by a mannose-resistant hemagglutination (MRHA) test with six species of erythrocytes and by a slide agglutination test with absorbed CFA/I or CFA/II antisera. The presence of CFAs was confirmed by immunodiffusion analyses done in Sweden. By these methods, it was found that 49 of 69 enterotoxin-producing E. coli strains isolated from patients carried CFA/I or CFA/II, whereas none of the nonenterotoxigenic E. coli isolates or the three toxin-positive strains isolated from healthy individuals carried these adhesins. All E. coli strains retained their MRHA ability after transportation to Sweden followed by one subculture and after storage at -70 degrees C (but not at room temperature) for 1 to 2 years without further subculturing. After 5 to 10 subcultures of the fresh isolates, however, 70% of the initially CFA/I- and 80% of the initially CFA/II-carrying strains analyzed did not hemagglutinate. The efficacy of different methods for detecting CFAs on the fresh isolates was compared with that of immunodiffusion. The sensitivity of MRHA with human blood group A erythrocytes for the detection of CFA/I was high (97%), but the specificity was only 69%. The sensitivity of MRHA with bovine erythrocytes for the detection of CFA/II in Bangladesh was very low but increased considerably when chicken erythrocytes were also used. Whereas both false-positive and false-negative reactions were obtained when absorbed CFA antisera were used for agglutination, antisera against purified CFAs were equally effective as immunodiffusion in identifying CFA/I and CFA/II-carrying strains.

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Selected References

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  1. Ahrén C. M., Svennerholm A. M. Synergistic protective effect of antibodies against Escherichia coli enterotoxin and colonization factor antigens. Infect Immun. 1982 Oct;38(1):74–79. doi: 10.1128/iai.38.1.74-79.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bäck E., Möllby R., Kaijser B., Stintzing G., Wadström T., Habte D. Enterotoxigenic Escherichia coli and other gram-negative bacteria of infantile diarrhea: surface antigens, hemagglutinins, colonization factor antigen, and loss of enterotoxigenicity. J Infect Dis. 1980 Sep;142(3):318–327. doi: 10.1093/infdis/142.3.318. [DOI] [PubMed] [Google Scholar]
  3. Cravioto A., Scotland S. M., Rowe B. Hemagglutination activity and colonization factor antigens I and II in enterotoxigenic and non-enterotoxigenic strains of Escherichia coli isolated from humans. Infect Immun. 1982 Apr;36(1):189–197. doi: 10.1128/iai.36.1.189-197.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Darfeuille A., Lafeuille B., Joly B., Cluzel R. A new colonization factor antigen (CFA/III) produced by enteropathogenic Escherichia coli O128:B12. Ann Microbiol (Paris) 1983 Jan-Feb;134A(1):53–64. doi: 10.1016/0769-2609(83)90103-5. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Evans D. G., Evans D. J., Jr, Tjoa W. S., DuPont H. L. Detection and characterization of colonization factor of enterotoxigenic Escherichia coli isolated from adults with diarrhea. Infect Immun. 1978 Feb;19(2):727–736. doi: 10.1128/iai.19.2.727-736.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Evans D. J., Jr, Evans D. G., DuPont H. L. Hemagglutination patterns of enterotoxigenic and enteropathogenic Escherichia coli determined with human, bovine, chicken, and guinea pig erythrocytes in the presence and absence of mannose. Infect Immun. 1979 Feb;23(2):336–346. doi: 10.1128/iai.23.2.336-346.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Giannella R. A. Suckling mouse model for detection of heat-stable Escherichia coli enterotoxin: characteristics of the model. Infect Immun. 1976 Jul;14(1):95–99. doi: 10.1128/iai.14.1.95-99.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Isaacson R. E., Richter P. Escherichia coli 987P pilus: purification and partial characterization. J Bacteriol. 1981 May;146(2):784–789. doi: 10.1128/jb.146.2.784-789.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Levine M. M., Ristaino P., Sack R. B., Kaper J. B., Orskov F., Orskov I. Colonization factor antigens I and II and type 1 somatic pili in enterotoxigenic Escherichia coli: relation to enterotoxin type. Infect Immun. 1983 Feb;39(2):889–897. doi: 10.1128/iai.39.2.889-897.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Merson M. H., Rowe B., Black R. E., Huq I., Gross R. J., Eusof A. Use of antisera for identification of enterotoxigenic Escherichia coli. Lancet. 1980 Aug 2;2(8188):222–224. doi: 10.1016/s0140-6736(80)90118-x. [DOI] [PubMed] [Google Scholar]
  15. Merson M. H., Sack R. B., Kibriya A. K., Al-Mahmood A., Adamed Q. S., Huq I. Use of colony pools for diagnosis of enterotoxigenic Escherichia coli diarrhea. J Clin Microbiol. 1979 Apr;9(4):493–497. doi: 10.1128/jcm.9.4.493-497.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Orskov F., Orskov I., Jann B., Jann K. Immunoelectrophoretic patterns of extracts from all Escherichia coli O and K antigen test strains: correlation with pathogenicity. Acta Pathol Microbiol Scand B Microbiol Immunol. 1971;79(2):142–152. doi: 10.1111/j.1699-0463.1971.tb02141.x. [DOI] [PubMed] [Google Scholar]
  17. Reis M. H., Matos D. P., de Castro A. F., Toledo M. R., Trabulsi L. R. Relationship among enterotoxigenic phenotypes, serotypes, and sources of strains in enterotoxigenic Escherichia coli. Infect Immun. 1980 Apr;28(1):24–27. doi: 10.1128/iai.28.1.24-27.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Stoll B. J., Rowe B., Glass R. I., Gross R. J., Huq I. Changes in serotypes of enterotoxigenic Escherichia coli in Dhaka over time: usefulness of polyvalent antisera. J Clin Microbiol. 1983 Oct;18(4):935–937. doi: 10.1128/jcm.18.4.935-937.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Svennerholm A. M., Wiklund G. Rapid GM1-enzyme-linked immunosorbent assay with visual reading for identification of Escherichia coli heat-labile enterotoxin. J Clin Microbiol. 1983 Apr;17(4):596–600. doi: 10.1128/jcm.17.4.596-600.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thomas L. V., Cravioto A., Scotland S. M., Rowe B. New fimbrial antigenic type (E8775) that may represent a colonization factor in enterotoxigenic Escherichia coli in humans. Infect Immun. 1982 Mar;35(3):1119–1124. doi: 10.1128/iai.35.3.1119-1124.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Willshaw G. A., Smith H. R., McConnell M. M., Barclay E. A., Krnjulac J., Rowe B. Genetic and molecular studies of plasmids coding for colonization factor antigen I and heat-stable enterotoxin in several escherichia coli serotypes. Infect Immun. 1982 Sep;37(3):858–868. doi: 10.1128/iai.37.3.858-868.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

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