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. 1982 Jul;49(1):239–246.

A new soluble antigen preparation of Toxoplasma gondii and its use in serological diagnosis.

H P Hughes, F Van Knapen, H J Atkinson, A H Balfour, D L Lee
PMCID: PMC1536634  PMID: 6751632

Abstract

A new method of solubilizing Toxoplasma gondii in 5% saponin and 5% octyl glucoside in combination (S-Ag) is described. When analysed by IEP, seven antigens could be detected. Four of these antigens (Ag 1, 2, 3 and 12) were shown by 125I-iodination to be of membrane origin. The remaining three (Ag 4, 5 and 6) were shown to be of intracellular origin and comprise the antigenic profile of more conventional preparations used in serodiagnosis. The S-Ag preparation was tested by ELISA (S-ELISA) in parallel with a freeze-thaw preparation containing only Ag 4, 5 and 6 (FT-ELISA), against the IgG-IFAT. The regression of the FT-ELISA against the IgG-IFAT was poor and there was a high incidence (28.9%) of false negative results. A second order polynomial provided a better fit and this predicted FT negative results on sera with IgG-IFAT titres lower than 1:1024. From this titre upwards, the test was specific but insensitive. The S-ELISA was more specific (5.6% false negatives) and more sensitive; polynomial terms could not provide a better fit. The improvement in the test using the S-Ag is considered to be due to the additional membrane antigens. The possible role of this solubilized antigen in future serodiagnosis is discussed.

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Selected References

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  1. Araujo F. G., Remington J. S. Antigenemia in recently acquired acute toxoplasmosis. J Infect Dis. 1980 Feb;141(2):144–150. doi: 10.1093/infdis/141.2.144. [DOI] [PubMed] [Google Scholar]
  2. BEVERLEY J. K. A., BEATTIE C. P. Standardization of the dye test for toxoplasmosis. J Clin Pathol. 1952 Nov;5(4):350–353. doi: 10.1136/jcp.5.4.350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carlier Y., Bout D., Dessaint J. P., Capron A., Van Knapen F., Ruitenberg E. J., Bergquist R., Huldt G. Evaluation of the enzyme-linked immunosorbent assay (ELISA) and other serological tests for the diagnosis of toxoplasmosis. Bull World Health Organ. 1980;58(1):99–105. [PMC free article] [PubMed] [Google Scholar]
  4. Hughes H. P., Balfour A. H. An investigation of the antigenic structure of Toxoplasma gondii. Parasite Immunol. 1981 Autumn;3(3):235–248. doi: 10.1111/j.1365-3024.1981.tb00403.x. [DOI] [PubMed] [Google Scholar]
  5. Hughes H. P. Characterization of the circulating antigen of Toxoplasma gondii. Immunol Lett. 1981 Jun;3(2):99–102. doi: 10.1016/0165-2478(81)90058-4. [DOI] [PubMed] [Google Scholar]
  6. Hunter D., Chadwick P., Balfour A. H., Bridges J. B. Examination of ovine foetal fluid for antibodies to Toxoplasma gondii by the dye test and an indirect immunofluorescence test specific for IgM. Br Vet J. 1982 Jan-Feb;138(1):29–34. doi: 10.1016/s0007-1935(17)31186-7. [DOI] [PubMed] [Google Scholar]
  7. JACOBS L., LUNDE M. N. A hemagglutination test for toxoplasmosis. J Parasitol. 1957 Jun;43(3):308–314. [PubMed] [Google Scholar]
  8. Karim K. A., Ludlam G. B. The relationship and significance of antibody titres as determined by various serological methods in glandular and ocular toxoplasmosis. J Clin Pathol. 1975 Jan;28(1):42–49. doi: 10.1136/jcp.28.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kramár J., Chalupský J., Hübner J. Relationship of the immunofluorescence reaction and the Sabin-Feldman reaction (second report). J Hyg Epidemiol Microbiol Immunol. 1970;14(2):129–134. [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. LUNDE M. N., JACOBS L. TOXOPLASMA HEMAGGLUTINATION AND DYE TEST ANTIBODIES IN EXPERIMENTALLY INFECTED RATS. J Parasitol. 1963 Dec;49:932–936. [PubMed] [Google Scholar]
  12. Pettersen E. K. Preparation of Toxoplasma gondii antigen for the complement fixation test. Acta Pathol Microbiol Scand. 1968;74(1):35–40. doi: 10.1111/j.1699-0463.1968.tb03452.x. [DOI] [PubMed] [Google Scholar]
  13. Phillips D. R., Morrison M. The arrangement of proteins in the human erythrocyte membrane. Biochem Biophys Res Commun. 1970 Jul 27;40(2):284–289. doi: 10.1016/0006-291x(70)91007-7. [DOI] [PubMed] [Google Scholar]
  14. Pokorný J. P., Frühbauer Z., Curík B., Zástera M. A Tween-ether preparation of Toxoplasma gondii antigen for the complement fixation test. Bull World Health Organ. 1972;46(1):127–130. [PMC free article] [PubMed] [Google Scholar]
  15. Pokorný J., Frühbauer Z., Curík B., Zástera M. Verification of tween-ether antigen for complement fixation test in the diagnostics of toxoplasmosis. J Hyg Epidemiol Microbiol Immunol. 1979;23(3):353–356. [PubMed] [Google Scholar]
  16. Sabin A. B., Feldman H. A. Dyes as Microchemical Indicators of a New Immunity Phenomenon Affecting a Protozoon Parasite (Toxoplasma). Science. 1948 Dec 10;108(2815):660–663. doi: 10.1126/science.108.2815.660. [DOI] [PubMed] [Google Scholar]
  17. Singh P., Chug T. D., Garg P. Toxoplasma IHA antibodies in human sera. Indian J Pathol Microbiol. 1978 Apr;21(2):125–129. [PubMed] [Google Scholar]
  18. Sulzer A. J., Hall E. C. Indirect fluorescent antibody tests for parasitic diseases. IV. Statistical study of variation in the indirect fluorescent antibody (IFA) test for toxoplasmosis. Am J Epidemiol. 1967 Sep;86(2):401–407. doi: 10.1093/oxfordjournals.aje.a120750. [DOI] [PubMed] [Google Scholar]
  19. Voller A., Bidwell D. E., Bartlett A., Fleck D. G., Perkins M., Oladehin B. A microplate enzyme-immunoassay for toxoplasma antibody. J Clin Pathol. 1976 Feb;29(2):150–153. doi: 10.1136/jcp.29.2.150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Waldeland H. Toxoplasmosis in sheep, the reliability of a microtiter system in Sabin and Feldman's dye test. Acta Vet Scand. 1976;17(4):426–431. doi: 10.1186/BF03547897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Walton B. C., Benchoff B. M., Brooks W. H. Comparison of the indirect fluorescent antibody test and methylene blue dye test for detection of antibodies to Toxoplasma gondii. Am J Trop Med Hyg. 1966 Mar;15(2):149–152. doi: 10.4269/ajtmh.1966.15.149. [DOI] [PubMed] [Google Scholar]
  22. van Knapen F., Panggabean S. O. Detection of circulating antigen during acute infections with Toxoplasma gondii by enzyme-linked immunosorbent assay. J Clin Microbiol. 1977 Dec;6(6):545–547. doi: 10.1128/jcm.6.6.545-547.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. van Knapen F., Panggabean S. O. Laboratory diagnosis of toxoplasmosis by means of an ELISA triple test (detection of class-specific IgG, IgM and circulating antigens). Antonie Van Leeuwenhoek. 1980;46(1):102–102. doi: 10.1007/BF00422239. [DOI] [PubMed] [Google Scholar]

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