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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1984 Mar;19(3):371–375. doi: 10.1128/jcm.19.3.371-375.1984

Rapid visual detection of Escherichia coli and Vibrio cholerae Heat-labile enterotoxins by nitrocellulose enzyme-linked immunosorbent assay.

L Beutin, L Bode, T Richter, G Peltre, R Stephan
PMCID: PMC271067  PMID: 6371043

Abstract

A modification of the enzyme-linked immunosorbent assay for a sensitive and rapid visual detection of heat-labile enterotoxins from Escherichia coli and Vibrio cholerae is described. Small amounts of bacterial supernatant fluids are bound to nitrocellulose filters which are used as sorbents in the nitrocellulose enzyme-linked immunosorbent assay. The test is based on the immunological similarity between V. cholerae and E. coli heat-labile enterotoxins. Six isolates of V. cholerae and 48 isolates of E. coli were examined for heat-labile enterotoxins by the nitrocellulose enzyme-linked immunosorbent assay and the Vero cell bioassay. With some strains, the nitrocellulose enzyme-linked immunosorbent assay was found to be more sensitive for detection of E. coli heat-labile enterotoxin than the Vero cell test. A similar result was obtained by endpoint titration of heat-labile enterotoxin-positive E. coli H10407 culture fluid in both assays. The sensitivity of the nitrocellulose enzyme-linked immunosorbent assay for the detection of purified cholera toxin was at a total level of 1 ng, which is a good result when compared with other serological assays.

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Selected References

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  1. Brill B. M., Wasilauskas B. L., Richardson S. H. Adaptation of the staphylococcal coagglutination technique for detection of heat-labile enterotoxin of Escherichia coli. J Clin Microbiol. 1979 Jan;9(1):49–55. doi: 10.1128/jcm.9.1.49-55.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clements J. D., Finkelstein R. A. Immunological cross-reactivity between a heat-labile enterotoxin(s) of Escherichia coli and subunits of Vibrio cholerae enterotoxin. Infect Immun. 1978 Sep;21(3):1036–1039. doi: 10.1128/iai.21.3.1036-1039.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DE S. N., CHATTERJE D. N. An experimental study of the mechanism of action of Vibriod cholerae on the intestinal mucous membrane. J Pathol Bacteriol. 1953 Oct;66(2):559–562. doi: 10.1002/path.1700660228. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Evans D. G., Evans D. J., Jr, Gorbach S. L. Identification of enterotoxigenic Escherichia coli and serum antitoxin activity by the vascular permeability factor assay. Infect Immun. 1973 Nov;8(5):731–735. doi: 10.1128/iai.8.5.731-735.1973. [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. J., Jr, Evans D. G. Inhibition of immune hemolysis: serological assay for the heat-labile enterotoxin of Excherichia coli. J Clin Microbiol. 1977 Jan;5(1):100–105. doi: 10.1128/jcm.5.1.100-105.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  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. Microtiter solid-phase radioimmunoassay for detection of Escherichia coli heat-labile enterotoxin. Infect Immun. 1977 Sep;17(3):541–545. doi: 10.1128/iai.17.3.541-545.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Guerrant R. L., Brunton L. L., Schnaitman T. C., Rebhun L. I., Gilman A. G. Cyclic adenosine monophosphate and alteration of Chinese hamster ovary cell morphology: a rapid, sensitive in vitro assay for the enterotoxins of Vibrio cholerae and Escherichia coli. Infect Immun. 1974 Aug;10(2):320–327. doi: 10.1128/iai.10.2.320-327.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gyles C. L. Immunological study of the heat-labile enterotoxins of Escherichia coli and Vibrio cholerae. Infect Immun. 1974 Mar;9(3):564–570. doi: 10.1128/iai.9.3.564-570.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Holmgren J., Svennerholm A. M. Enzyme-linked immunosorbent assays for cholera serology. Infect Immun. 1973 May;7(5):759–763. doi: 10.1128/iai.7.5.759-763.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Honda T., Samakoses R., Sornchai C., Takeda Y., Miwatani T. Detection by a staphylococcal coagglutination test of heat-labile enterotoxin-producing enterotoxigenic Escherichia coli. J Clin Microbiol. 1983 Apr;17(4):592–595. doi: 10.1128/jcm.17.4.592-595.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Honda T., Taga S., Takeda Y., Miwatani T. Modified Elek test for detection of heat-labile enterotoxin of enterotoxigenic Escherichia coli. J Clin Microbiol. 1981 Jan;13(1):1–5. doi: 10.1128/jcm.13.1.1-5.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ito T., Kuwahara S., Yokota T. Automatic and manual latex agglutination tests for measurement of cholera toxin and heat-labile enterotoxin of Escherichia coli. J Clin Microbiol. 1983 Jan;17(1):7–12. doi: 10.1128/jcm.17.1.7-12.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Moon H. W., Whipp S. C., Engstrom G. W., Baetz A. L. Response of the rabbit ileal loop to cell-free products from Escherichia coli enteropathogenic for swine. J Infect Dis. 1970 Feb;121(2):182–187. doi: 10.1093/infdis/121.2.182. [DOI] [PubMed] [Google Scholar]
  16. O'Connor C. G., Ashman L. K. Application of the nitrocellulose transfer technique and alkaline phosphatase conjugated anti-immunoglobulin for determination of the specificity of monoclonal antibodies to protein mixtures. J Immunol Methods. 1982 Oct 29;54(2):267–271. doi: 10.1016/0022-1759(82)90068-0. [DOI] [PubMed] [Google Scholar]
  17. Palfree R. G., Elliott B. E. An enzyme-linked immunosorbent assay (ELISA) for detergent solubilized Ia glycoproteins using nitrocellulose membrane discs. J Immunol Methods. 1982 Aug 13;52(3):395–408. doi: 10.1016/0022-1759(82)90010-2. [DOI] [PubMed] [Google Scholar]
  18. Sack D. A., Sack R. B. Test for enterotoxigenic Escherichia coli using Y-1 adrenal cells in miniculture. Infect Immun. 1975 Feb;11(2):334–336. doi: 10.1128/iai.11.2.334-336.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sack R. B., Froehlich J. L. Antigenic similarity of heat-labile enterotoxins from diverse strains of Escherichia coli. J Clin Microbiol. 1977 Jun;5(6):570–572. doi: 10.1128/jcm.5.6.570-572.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Salonen E. M., Vaheri A. Immobilization of viral and mycoplasma antigens and of immunoglobulins on polystyrene surface for immunoassays. J Immunol Methods. 1979;30(3):209–218. doi: 10.1016/0022-1759(79)90095-4. [DOI] [PubMed] [Google Scholar]
  21. Serafim M. B., de Castro A. F., Leonardo M. B., Monteiro A. R. Single radial immune hemolysis test for detection of Escherichia coli thermolabile enterotoxin. J Clin Microbiol. 1981 Nov;14(5):473–478. doi: 10.1128/jcm.14.5.473-478.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Shah D. B., Kauffman P. E., Boutin B. K., Johnson C. H. Detection of heat-labile-enterotoxin-producing colonies of Escherichia coli and Vibrio cholerae by solid-phase sandwich radioimmunoassays. J Clin Microbiol. 1982 Sep;16(3):504–508. doi: 10.1128/jcm.16.3.504-508.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  24. Speirs J. I., Stavric S., Konowalchuk J. Assay of Escherichia coli heat-labile enterotoxin with vero cells. Infect Immun. 1977 May;16(2):617–622. doi: 10.1128/iai.16.2.617-622.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yolken R. H., Greenberg H. B., Merson M. H., Sack R. B., Kapikian A. Z. Enzyme-linked immunosorbent assay for detection of Escherichia coli heat-labile enterotoxin. J Clin Microbiol. 1977 Nov;6(5):439–444. doi: 10.1128/jcm.6.5.439-444.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. de Savigny D., Voller A. The communication of ELISA data from laboratory to clinician. J Immunoassay. 1980;1(1):105–128. doi: 10.1080/01971528008055779. [DOI] [PubMed] [Google Scholar]

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