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. 1985 Dec;22(6):893–896. doi: 10.1128/jcm.22.6.893-896.1985

Evaluation of a competitive enzyme-linked immunosorbent assay for porcine Escherichia coli heat-stable enterotoxin.

B Rönnberg, O Söderlind, T Wadström
PMCID: PMC271846  PMID: 3905848

Abstract

A competitive enzyme-linked immunosorbent assay (ELISA) was compared with the conventional suckling mouse assay for the identification of heat-stable enterotoxin (STa) in samples from piglets suffering from diarrhea. A total of 110 Escherichia coli isolates, 22 primary cultures, and 26 fecal samples from piglets up to 8 weeks of age with diarrhea were compared in parallel by both assays. Of the 110 isolates tested, all gave consistent results by the ELISA and the suckling mouse assay; 60 strains were negative and 50 strains produced STa by both tests. Identical results were obtained when 22 primary agar cultures were screened for STa production by both methods; 6 were found to produce STa, while 16 did not. When 26 fecal samples were tested for the presence of STa, 10 were negative and 12 were positive by both assays. One of the remaining four samples gave questionable positive results by both the suckling mouse assay and the ELISA, but E. coli isolated from this sample gave positive results by both tests. The remaining three samples were negative by the suckling mouse assay, but gave questionable positive results by the ELISA. E. coli isolates from these three samples were always negative by both assays. The ELISA used in this study provides a reliable and convenient method for diagnosing STa-producing enterotoxigenic E. coli of porcine origin.

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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. 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]
  3. 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]
  4. Frantz J. C., Robertson D. C. Immunological properties of Escherichia coli heat-stable enterotoxins: development of a radioimmunoassay specific for heat-stable enterotoxins with suckling mouse activity. Infect Immun. 1981 Jul;33(1):193–198. doi: 10.1128/iai.33.1.193-198.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Johnson W. M., Lior H., Johnson K. G. Heat-stable enterotoxin from Escherichia coli: factors involved in growth and toxin production. Infect Immun. 1978 May;20(2):352–359. doi: 10.1128/iai.20.2.352-359.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Klipstein F. A., Engert R. F., Houghten R. A., Rowe B. Enzyme-linked immunosorbent assay for Escherichia coli heat-stable enterotoxin. J Clin Microbiol. 1984 Jun;19(6):798–803. doi: 10.1128/jcm.19.6.798-803.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lallier R., Bernard F., Gendreau M., Lazure C., Seidah N. G., Chrétien M., St-Pierre S. A. Isolation and purification of Escherichia coli heat-stable enterotoxin of porcine origin. Anal Biochem. 1982 Dec;127(2):267–275. doi: 10.1016/0003-2697(82)90171-3. [DOI] [PubMed] [Google Scholar]
  10. Lockwood D. E., Robertson D. C. Development of a competitive enzyme-linked immunosorbent assay (ELISA) for Escherichia coli heat-stable enterotoxin (STa). J Immunol Methods. 1984 Dec 31;75(2):295–307. doi: 10.1016/0022-1759(84)90113-3. [DOI] [PubMed] [Google Scholar]
  11. Moseley S. L., Echeverria P., Seriwatana J., Tirapat C., Chaicumpa W., Sakuldaipeara T., Falkow S. Identification of enterotoxigenic Escherichia coli by colony hybridization using three enterotoxin gene probes. J Infect Dis. 1982 Jun;145(6):863–869. doi: 10.1093/infdis/145.6.863. [DOI] [PubMed] [Google Scholar]
  12. Murray B. E., Seriwatana J., Echeverria P. Toxin detection after storage or cultivation of enterotoxigenic with colicinogenic Escherichia coli: a possible mechanism for toxin-negative pools. J Clin Microbiol. 1981 Jan;13(1):179–183. doi: 10.1128/jcm.13.1.179-183.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Rönnberg B., Wadström T. Improved method for purification of human Escherichia coli heat-stable enterotoxin by hydrophobic interaction, molecular-sieve and high performance ion exchange chromatography. Prep Biochem. 1983;13(3):245–260. doi: 10.1080/00327488308064251. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. So M., McCarthy B. J. Nucleotide sequence of the bacterial transposon Tn1681 encoding a heat-stable (ST) toxin and its identification in enterotoxigenic Escherichia coli strains. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4011–4015. doi: 10.1073/pnas.77.7.4011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Thompson M. R., Brandwein H., LaBine-Racke M., Giannella R. A. Simple and reliable enzyme-linked immunosorbent assay with monoclonal antibodies for detection of Escherichia coli heat-stable enterotoxins. J Clin Microbiol. 1984 Jul;20(1):59–64. doi: 10.1128/jcm.20.1.59-64.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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