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. 1984 Jul;20(1):59–64. doi: 10.1128/jcm.20.1.59-64.1984

Simple and reliable enzyme-linked immunosorbent assay with monoclonal antibodies for detection of Escherichia coli heat-stable enterotoxins.

M R Thompson, H Brandwein, M LaBine-Racke, R A Giannella
PMCID: PMC271246  PMID: 6540277

Abstract

We have developed a sensitive and specific competitive enzyme-linked immunosorbent assay for Escherichia coli heat-stable enterotoxins consisting of methanol-soluble, suckling mouse active peptides with similar core sequences (STa) by using monoclonal antibodies prepared against STa purified from a human isolate. The assay can detect 3 to 20 pg of purified STa, depending on the monoclonal antibody used in the assay. The assay is rapid, requiring ca. 1 h to complete. With this competitive enzyme-linked immunosorbent assay, we measured STa production by enterotoxigenic E. coli directly in Casamino Acid-yeast extract culture supernatants. The assay was suitable for measuring STa in culture supernatants from human, bovine, and porcine E. coli isolates. No cross-reactivity was observed with heat-labile enterotoxin, cholera toxin, or heat-stable enterotoxin STb, which is a methanol-insoluble peptide(s) active in the ligated pig jejunal loop test. A 100% correlation of toxin production was found by comparing the enzyme-linked immunosorbent assay with the previously established radioimmunoassay for STa and with suckling mouse activity.

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Selected References

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  1. Aimoto S., Takao T., Shimonishi Y., Hara S., Takeda T., Takeda Y., Miwatani T. Amino-acid sequence of a heat-stable enterotoxin produced by human enterotoxigenic Escherichia coli. Eur J Biochem. 1982 Dec 15;129(2):257–263. doi: 10.1111/j.1432-1033.1982.tb07047.x. [DOI] [PubMed] [Google Scholar]
  2. Burgess M. N., Bywater R. J., Cowley C. M., Mullan N. A., Newsome P. M. Biological evaluation of a methanol-soluble, heat-stable Escherichia coli enterotoxin in infant mice, pigs, rabbits, and calves. Infect Immun. 1978 Aug;21(2):526–531. doi: 10.1128/iai.21.2.526-531.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chan S. K., Giannella R. A. Amino acid sequence of heat-stable enterotoxin produced by Escherichia coli pathogenic for man. J Biol Chem. 1981 Aug 10;256(15):7744–7746. [PubMed] [Google Scholar]
  4. Clements J. D., Finkelstein R. A. Isolation and characterization of homogeneous heat-labile enterotoxins with high specific activity from Escherichia coli cultures. Infect Immun. 1979 Jun;24(3):760–769. doi: 10.1128/iai.24.3.760-769.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dreyfus L. A., Frantz J. C., Robertson D. C. Chemical properties of heat-stable enterotoxins produced by enterotoxigenic Escherichia coli of different host origins. Infect Immun. 1983 Nov;42(2):539–548. doi: 10.1128/iai.42.2.539-548.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Giannella R. A., Drake K. W., Luttrell M. Development of a radioimmunoassay for Escherichia coli heat-stable enterotoxin: comparison with the suckling mouse bioassay. Infect Immun. 1981 Jul;33(1):186–192. doi: 10.1128/iai.33.1.186-192.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Kauffman P. E. Production and evaluation of antibody to the heat-stable enterotoxin from a human strain of enterotoxigenic Escherichia coli. Appl Environ Microbiol. 1981 Oct;42(4):611–614. doi: 10.1128/aem.42.4.611-614.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lazure C., Seidah N. G., Chrétien M., Lallier R., St-Pierre S. Primary structure determination of Escherichia coli heat-stable enterotoxin of porcine origin. Can J Biochem Cell Biol. 1983 May;61(5):287–292. doi: 10.1139/o83-039. [DOI] [PubMed] [Google Scholar]
  11. Moon H. W. Pathogenesis of enteric diseases caused by Escherichia coli. Adv Vet Sci Comp Med. 1974;18(0):179–211. [PubMed] [Google Scholar]
  12. Moseley S. L., Hardy J. W., Hug M. I., Echeverria P., Falkow S. Isolation and nucleotide sequence determination of a gene encoding a heat-stable enterotoxin of Escherichia coli. Infect Immun. 1983 Mar;39(3):1167–1174. doi: 10.1128/iai.39.3.1167-1174.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Rönnberg B., Wadström T., Jörnvall H. Structure of a heat-stable enterotoxin produced by a human strain of Escherichia coli. Differences from the toxin of another human strain suggest the presence of compensated amino acid exchanges. FEBS Lett. 1983 May 8;155(2):183–186. doi: 10.1016/0014-5793(82)80598-x. [DOI] [PubMed] [Google Scholar]
  14. Sack R. B. Human diarrheal disease caused by enterotoxigenic Escherichia coli. Annu Rev Microbiol. 1975;29:333–353. doi: 10.1146/annurev.mi.29.100175.002001. [DOI] [PubMed] [Google Scholar]
  15. Saeed A. M., Sriranganathan N., Cosand W., Burger D. Purification and characterization of heat-stable enterotoxin from bovine enterotoxigenic Escherichia coli. Infect Immun. 1983 May;40(2):701–707. doi: 10.1128/iai.40.2.701-707.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Smith H. W., Gyles C. L. The relationship between two apparently different enterotoxins produced by enteropathogenic strains of Escherichia coli of porcine origin. J Med Microbiol. 1970 Aug;3(3):387–401. doi: 10.1099/00222615-3-3-387. [DOI] [PubMed] [Google Scholar]
  17. Staples S. J., Asher S. E., Giannella R. A. Purification and characterization of heat-stable enterotoxin produced by a strain of E. coli pathogenic for man. J Biol Chem. 1980 May 25;255(10):4716–4721. [PubMed] [Google Scholar]

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