Current Microbiology

, Volume 1, Issue 1, pp 19–23 | Cite as

Identification ofEscherichia coli heat-labile enterotoxin by means of a ganglioside immunosorbent assay (GM1-ELISA) procedure

  • Ann-Mari Svennerholm
  • Jan Holmgren
Article

Abstract

A sensitive, quantitative method for determination ofEscherichia coli heat-labile enterotoxin (LT) is presented. The assay is based on the specific binding of LT to polystyrene-adsorbed GM1 ganglioside and subsequent enzyme immunological demonstration of the bound toxin. Enterotoxin titers determined with this GM1 enzyme-linked immunosorbent assay (ELISA) method agreed closely with those obtained with the adrenal cell bioassay. The GM1-ELISA procedure was capable of demonstrating LT in allE. coli overnight cultures that gave positive adrenal cell results. The simplicity and high reproducibility of the described method should make it well suited for routine laboratory diagnosis of LT enterotoxigenicE. coli strains.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Literature Cited

  1. 1.
    Craig, J. P. 1965. A permeability factor (toxin) found in cholera stools and culture filtrates and its neutralization by convalescent cholera sera. Nature207:614–616.PubMedCrossRefGoogle Scholar
  2. 2.
    Dafni, Z., Robbins, J. B. 1976. Purification of heat-labile enterotoxin fromEscherichia coli 078:H11 by affinity chromatography with antisera toVibrio cholerae toxin. Journal of Infectious Diseases133:S138-S141.Google Scholar
  3. 3.
    Donta, S. T., Moon, H. W., Shipp, S. C. 1974. Detection of heat-labileEscherichia coli enterotoxin with the use of adrenal cells in tissue culture. Science183:334–336.PubMedCrossRefGoogle Scholar
  4. 4.
    Engvall, E. 1977. Quantitative enzyme immunoassay (ELISA) in microbiology. Medical Biology55:193–200.PubMedGoogle Scholar
  5. 5.
    Engvall, E., Perlman, P. 1972. Enzyme-linked immunosorbent assay, ELISA. III. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. Journal of Immunology109:129–135.Google Scholar
  6. 6.
    Evans, D. J., Jr., Evans, D. G. 1977. Direct serological assay for the heat-labile enterotoxin ofEscherichia coli using passive immunohemolysis. Infection and Immunity16:604–609.PubMedGoogle Scholar
  7. 7.
    Evans, D. J., Jr., Evans, D. G., Richarson, S. H., Gorbach, S. L. 1976. Purification of the polymyxin-released, heat-labile enterotoxin ofEscherichia coli. Journal of Infectious Diseases133:S97-S102.Google Scholar
  8. 8.
    Greenberg, H. B., Sack, D. A., Rodriguez, W., Sack, R. B., Wyatt, R. G., Kalica, A. R., Horswood, R. L., Chanock, R. M., Kapikian, A. Z. 1977. Microtier solid-phase radioimmunoassay for detection ofEscherichia coli heatlabile enterotoxin. Infection and Immunity17:541–545.PubMedGoogle Scholar
  9. 9.
    Guerrant, R. L., Branton, L. L., Schartman, T. C., Rebhun, L. I., Gilman, A. G. 1974. Cyclic adenosine monophosphate and alteration of the Chinese hamster ovary cell morphology: A rapid sensitivein vitro assay for enterotoxins ofVibrio cholerae andEscherichia coli. Infection and Immunity10:320–327.PubMedGoogle Scholar
  10. 10.
    Holmgren, J. 1973. Comparison of the tissue receptors forVibrio cholerae andEscherichia coli enterotoxins by means of gangliosides and natural cholera toxoid. Infection and Immunity8:851–859.PubMedGoogle Scholar
  11. 11.
    Holmgren, J., Svennerholm, A.-M. 1973. Enzyme-linked immunosorbent assay for cholera serology. Infection and Immunity7:759–763.PubMedGoogle Scholar
  12. 12.
    Holmgren, J., Svennerholm, A.-M., Lönnroth, I., Fall-Persson, M., Markman, B., Lundbäck, H. 1977. Development of improved cholera vaccine based on subunit toxoid. Nature269:602–604.PubMedCrossRefGoogle Scholar
  13. 13.
    Holmgren, J., Söderlind, O., Wadström, T. 1973. Cross-reactivity between heat labile enterotoxins ofVibrio cholera andEscherichia coli in neutralization tests in rabbit ileum and skin. Acta Pathologica et Microbiologica Scandinavica Section B81:757–762.Google Scholar
  14. 14.
    Sack, R. B. 1975. Human diarrheal disease caused by enterotoxigenicEscherichia coli. Annual Review of Microbiology26:333–353.CrossRefGoogle Scholar
  15. 15.
    Short, L. C., Jr., Kurtz, H. J. 1971. Properties of hemolytic activities ofEscherichia coli. Infection and Immunity3:678–687.PubMedGoogle Scholar
  16. 16.
    Smith, H. W. 1963. The hemolysins ofE. coli. Journal of Pathology and Bacteriology85:197–211.PubMedCrossRefGoogle Scholar
  17. 17.
    Svennerholm, L. 1972. Gangliosides, isolation, pp. 464–474. In: Whistler, R. L., BeMiller, J. N. (eds.), Methods in carbohydrate chemistry, vol. 6. New York: Academic Press.Google Scholar

Copyright information

© Springer-Verlag New York Inc 1978

Authors and Affiliations

  • Ann-Mari Svennerholm
    • 1
  • Jan Holmgren
    • 1
  1. 1.Institute of Medical MicrobiologyUniversity of GöteborgGöteborgSweden

Personalised recommendations