Abstract
Automated and manual latex agglutination methods were employed to measure cholera toxin (CT), heat-labile enterotoxin (LT) of Escherichia coli, and their subunits A and B. Dow polystyrene latex particles (diameter, 0.22 microns) and polystyrene-chlorostyrene latex particles (diameter, 1 micron) were sensitized by rabbit-specific immunoglobulin for each antigen and used as the reagents of the automated and manual agglutination tests, respectively. Automated agglutination was performed by a nephelometric assay system measuring time-dependent differences of light scattering due to agglutination, and manual latex agglutination was carried out in microtiter plates. As low as 1,000 and 31 pg of CT per ml were estimated by the automated and manual agglutination tests, respectively. Using these methods, the amount of CT and LT was measured in several clinical isolates of Vibrio cholerae and E. coli. Furthermore, it was discovered that cyclic AMP is not essential for the production of CT by measuring the amount of the toxin in numbers of cyclic AMP-dependent mutants of V. cholerae (with the agglutination tests).
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Selected References
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- Blume P., Greenberg L. J. Application of differential light scattering to the latex agglutination assay for rheumatoid factor. Clin Chem. 1975 Aug;21(9):1234–1237. [PubMed] [Google Scholar]
- Clements J. D., Finkelstein R. A. Demonstration of shared and unique immunological determinants in enterotoxins from Vibrio cholerae and Escherichia coli. Infect Immun. 1978 Dec;22(3):709–713. doi: 10.1128/iai.22.3.709-713.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Craig J. P. A permeability factor (toxin) found in cholera stools and culture filtrates and its neutralization by convalescent cholera sera. Nature. 1965 Aug 7;207(997):614–616. doi: 10.1038/207614a0. [DOI] [PubMed] [Google Scholar]
- Donta S. T., Moon H. W., Whipp S. C. Detection of heat-labile Escherichia coli enterotoxin with the use of adrenal cells in tissue culture. Science. 1974 Jan 25;183(4122):334–336. doi: 10.1126/science.183.4122.334. [DOI] [PubMed] [Google Scholar]
- Finkelstein R. A., LaRue M. K., Johnston D. W., Vasil M. L., Cho G. J., Jones J. R. Isolation and properties of heat-labile enterotoxin(s) from enterotoxigenic Escherichia coli. J Infect Dis. 1976 Mar;133 (Suppl):120–137. doi: 10.1093/infdis/133.supplement_1.s120. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Guerrant R. L., Moore R. A., Kirschenfeld P. M., Sande M. A. Role of toxigenic and invasive bacteria in acute diarrhea of childhood. N Engl J Med. 1975 Sep 18;293(12):567–572. doi: 10.1056/NEJM197509182931201. [DOI] [PubMed] [Google Scholar]
- 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]
- Ohtomo N., Muraoka T., Tashiro A., Zinnaka Y., Amako K. Size and structure of the cholera toxin molecule and its subunits. J Infect Dis. 1976 Mar;133 (Suppl):31–40. doi: 10.1093/infdis/133.supplement_1.s31. [DOI] [PubMed] [Google Scholar]
- PLOTZ C. M., SINGER J. M. The latex fixation test. I. Application to the serologic diagnosis of rheumatoid arthritis. Am J Med. 1956 Dec;21(6):888–892. [PubMed] [Google Scholar]
- Quash G., Roch A. M., Niveleau A., Grange J., Keolouangkhot T., Huppert J. The preparation of latex particles with covalently bound polyamines, IgG and measles agglutinins and their use in visual agglutination tests. J Immunol Methods. 1978;22(1-2):165–174. doi: 10.1016/0022-1759(78)90069-8. [DOI] [PubMed] [Google Scholar]
- Tsukamoto T., Kinoshita Y., Taga S., Takeda Y., Miwatani T. Value of passive immune hemolysis for detection of heat-labile enterotoxin produced by enterotoxigenic Escherichia coli. J Clin Microbiol. 1980 Dec;12(6):768–771. doi: 10.1128/jcm.12.6.768-771.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yokota T., Kuwahara S. Adenosine 3',5'-cyclic monophosphate-deficient mutants of Vibrio cholerae. J Bacteriol. 1974 Oct;120(1):106–113. doi: 10.1128/jb.120.1.106-113.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- van Heyningen S. The subunits of cholera toxin: structure, stoichiometry, and function. J Infect Dis. 1976 Mar;133 (Suppl):5–13. doi: 10.1093/infdis/133.supplement_1.s5. [DOI] [PubMed] [Google Scholar]