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Assay for heat-labile Escherichia coli enterotoxin using sandwich erythroimmunoassay

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Abstract

We investigated the possibility of using a sheep erythrocyte-antibody conjugate as reagent in a sandwich erythroimmunoassay (SERIA) procedure to detect and titrate Escherichia coli heat-labile enterotoxin (LT) with the naked eye. In this assay, which is based on the immunological similarity between Vibrio cholerae toxin (CT) and LT, rabbit anti-CT IgG was used as immunosorbent, and sheep erythrocytes, sensitized with the rabbit anti-CT antibodies, were used as indicator. The sensitivity of the test was demonstrated by a comparative study using an enzyme-linked immunosorbent assay (ELISA). The results obtained by SERIA with 130 samples correlated well with those of a Vero cell assay and a GM1-ELISA. The test is easy, relatively cheap and as sensitive as other standard techniques; it is particularly suited for field laboratories, especially in tropical countries, and a large number of strains may be examined daily.

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References

  1. Beuthin L, Bode L, Richter T, Peltre G, Stephan R (1984) Rapid visual detection of Escherichia coli and Vibrio cholerae heat labile enterotoxins by nitrocellulose enzyme-linked immunosorbent assay. J Clin Microbiol 19:371–375

    Google Scholar 

  2. Brill BM, Wasilanskas BL, Richardson SH (1979) Adaptation of the staphylococcal coagglutination technique for detection of heat-labile enterotoxin of Escherichia coli. J Clin Microbiol 9:49–55

    Google Scholar 

  3. Burgess MN, Bywater RJ, Cowley CM, Mullan NA, Newsome PM (1978) Biological evaluation of a methanol-soluble, heat-stable Escherichia coli enterotoxin in infant mice, pigs, rabbits, and calves. Infect Immun 21:526–531

    Google Scholar 

  4. Caprioli A, Falbo V, Roda LG, Ruggeri FM, Zona C (1983) Partial purification and characterization of an Escherichia coli toxic factor that induces morphological cell alterations. Infect Immun 39:1300–1305

    CAS  PubMed  Google Scholar 

  5. Chan S, Giannella RA (1981) Amino acid sequence of heat stable enterotoxin produced by Escherichia coli pathogenic for man. J Biol Chem 256:7744–7746

    Google Scholar 

  6. Clements JD, Finkelstein RA (1979) Isolation and characterization of homogeneous heat-labile enterotoxins with high specific activity from Escherichia coli cultures. Infect Immun 24:760–769

    Google Scholar 

  7. Craig JP (1965) A permeability factor (toxin) found in cholera stool and culture filtrates and its neutralisation by convalescent sera. Nature 207:614–616

    Google Scholar 

  8. Dallas WS, Falkow S (1980) Amino acid sequence homology between cholera toxin and Escherichia coli heat-labile toxin. Nature 288:499–501

    Google Scholar 

  9. Donta ST, Moon HW, Whipp SC (1974) Detection of heat-labile Escherichia coli enterotoxin with the use of adrenal cells in tissue cultures. Science 183:334–336

    Google Scholar 

  10. Evans DJ, Evans G, Dupont HL, Orskov F, Orskov I (1977) Patterns of loss of enterotoxigenicity by Eschericia coli isolated from adults with diarrhea: suggestive evidence for interelationship with serotype. Infect Immun 17:105–111

    Google Scholar 

  11. Finkelstein RA, Yang Z (1983) Rapid test for identification of heat-labile enterotoxin producing Escherichia coli colonies. J Clin Microbiol 18:23–28

    Google Scholar 

  12. Germani Y, Begaud E, Dassy B, Legonidec G, Guesdon JL (1984) Method for the detection and dosage of the Escherichia coli heat-labile enterotoxin by an enzyme-linked immunosorbent assay using a new simple kit test. Ann Microbiol Inst Pasteur 135:296–310

    Google Scholar 

  13. Germani Y, Dassy B, Legonidec G, Guesdon JL (1984) Titration of rabbit anti-Escherichia coli heat-labile enterotoxin antibodies by an enzyme-linked immunosorbent assay. Ann Immunol Inst Pasteur 135:251–260

    Google Scholar 

  14. Germani Y, Montaville B, Fauran C, Brethes B (1985) Survey in Vanuatu on enterotoxigeni Escherichia coli in children and infants with and without acute diarrhea. J Clin Microbiol 21:630–633

    Google Scholar 

  15. Green BA, Neill RJ, Ruyechan WT, Holmes RK (1983) Evidence that a new enterotoxin of Escherichia coli which adenylate cyclase in eucaryotic target cells is not plasmid mediated. Infect Immun 41:383–390

    Google Scholar 

  16. Greenberg HD, Sack DA, Rodriguez W, Sack RB, Wyatt RG, Kalica AR, Harswood RI, Chanock RM, Kapikian AZ (1977) Microtiter solid-phase radioimmunoassay for detection of Escherichia coli heat-labile enterotoxin. Infect Immun 17:541–545

    Google Scholar 

  17. Greenberg RN, Guerrant RL (1980) E. coli heat stable enterotoxin. Pharmacol Ther 11:507–531

    Google Scholar 

  18. Guerrant RL, Brunton LL, Schnaitman TC, Rebhrun LI, Gilman AG (1974) Cyclic adenosine monophosphate and alteration of the chinese hamster ovary cell morphology: a rapid sensitive in vitro assay for enterotoxins of Vibrio cholerae and Escherichia coli. Infect Immun 10:320–327

    Google Scholar 

  19. Guesdon JL, Avrameas S (1980) Sensitive titration of antibodies and antigens using erythroimmunoassay. Ann Immunol Inst Pasteur 131:389–396

    Google Scholar 

  20. Gupta SK, Guesdon JL, Avrameas S, Talwar GP (1985) Solid-phase competitive and sandwich-type erythroimmunoassay for human chorionic gonadotropin. J Immun Meth 80:177–188

    Google Scholar 

  21. Honda T, Taga S, Takeda Y and Miwatani T (1981) Modified Elek test for detection of heat-labile enterotoxin of enterotoxigenic Escherichia coli. J Clin Microbiol 13:1–5

    Google Scholar 

  22. Honda T, Takeda Y, Miwatani T (1981) Isolation of special antibodies which react only with homologous enterotoxins from Vibrio cholerae and enterotoxigenic Escherichia coli. Infect Immun 34:333–336

    Google Scholar 

  23. Konowalchuck J, Speirs JI, Stavric S (1977) Vero response to a cytotoxin of Escherichia coli. Infect Immun 18:775–779

    Google Scholar 

  24. Merson MH, Morris GK, Sack DA, Wells JG, Feeley JC, Sack RB, Creek WB, Kapikian AZ, Gangarosa EJ (1976) Traveler's diarrhea in Mexico: a prospective study of physicians and family members attending a congress. N Engl J Med 294:1299–1305

    Google Scholar 

  25. Moon HW, Whipp SC, Engström GW, Baelz AL (1970) Response of rabbit ileal loop to cellfree products from Escherichia coli enteropathogenic for swine. J Infect Dis 128:182–187

    Google Scholar 

  26. Mundell DH, Anselmo CR, Wishnow RM (1976) Factors influencing heat-labile Escherichia coli enterotoxin activity. Infect Immun 14:383–388

    Google Scholar 

  27. Ristaino PA, Levine MM, Young CR (1983) Improved GM1-enzyme-linked immunosorbent assay for detection of Escherichia coli heat-labile enterotoxin. J Clin Microbiol 18:808–815

    Google Scholar 

  28. Serafim MB, Pestana de Castro AF, Leonardo MB, Monteiro AR (1981) Single radial immune hemolysis test for detection of Escherichia coli thermolabile enterotoxin. J Clin Microbiol 14:473–478

    Google Scholar 

  29. Snyder JD, Merson MH (1982) The magnitude of the global problem of acute diarrhoeal disease: a review of active surveillance data. Bull WHO 60:605–613

    Google Scholar 

  30. Speirs JI, Stavric S, Konowalchuck J (1977) Assay of Escherichia coli heat-labile enterotoxin with Vero cells. Infect Immun 16:617–622

    Google Scholar 

  31. Svennerholm AM, Holmgren J (1978) Identification of Escherichia coli heat-labile enterotoxin by means of a ganglioside immunosorbent assay (GM1-ELISA) procedure. Curr Microbiol 1:19–23

    Google Scholar 

  32. Yano T, Oliveira MS, Fontes CF, de Almeida ACP, de Castro AFP (1982) Detection of heat-labile (LT) enterotoxin of enterotoxigenic Escherichia coli by radial immune hemolysis test: a modification for clinical use. Med Microbiol Immunol 171:171–178

    Google Scholar 

  33. Yolken BH, Greenberg HB, Merson MH, Sack RB, Kapikian AZ (1977) Enzyme linked immunosorbent assay for detection of Escherichia coli heat-labile enterotoxin. J Clin Microbiol 6:439–444

    Google Scholar 

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Authors and Affiliations

  1. Unité de Bactériologie Expérimentale, Institut Pasteur, BP 61, Nouméa, New Caledonia

    Y. Germani & E. Begaud

  2. Laboratoire des Sondes Froides, Institut Pasteur, 28 rue du Dr. Roux, F-75 724, Paris Cedex 15, France

    J. L. Guesdon

Authors
  1. Y. Germani
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  2. E. Begaud
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  3. J. L. Guesdon
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Germani, Y., Begaud, E. & Guesdon, J.L. Assay for heat-labile Escherichia coli enterotoxin using sandwich erythroimmunoassay. Med Microbiol Immunol 176, 89–97 (1987). https://doi.org/10.1007/BF00200680

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  • DOI: https://doi.org/10.1007/BF00200680

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