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. 1985 Nov;62(2):262–269.

Serodiagnosis of toxoplasma infection using a purified parasite protein (P30).

F Santoro, D Afchain, R Pierce, J Y Cesbron, G Ovlaque, A Capron
PMCID: PMC1577441  PMID: 2417762

Abstract

The major surface protein (P30) of Toxoplasma gondii has been purified by immunoabsorption with anti-P30 monoclonal antibodies linked to a glutardialdehyde activated affinity absorbant. SDS-PAGE analysis of the eluted material followed by silver staining showed only a single band of 30,000 mol wt. Western blotting using antibodies from a rabbit immunized with purified P30 against the total Toxoplasma extract separated by SDS-PAGE again revealed an unique antigen of 30,000 daltons. The presence of repeated epitopes within P30 was confirmed by a two-site/one-antibody radiometric assay with the purified protein. Sandwich ELISA procedures with purified P30 clearly demonstrated that all 37 tested patients with acute toxoplasmosis presented significantly high levels of IgM anti-P30 antibodies. In addition, all 40 tested patients with chronic toxoplasma infection also showed high IgG anti-P30 antibody levels. These findings represent an essential step for the development of new reagents for the diagnosis of toxoplasmosis.

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Selected References

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  1. Araujo F. G., Remington J. S. Partially purified antigen preparations of Toxoplasma gondii protect against lethal infection in mice. Infect Immun. 1984 Jul;45(1):122–126. doi: 10.1128/iai.45.1.122-126.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Camargo M. E., Ferreira A. W., Mineo J. R., Takiguti C. K., Nakahara O. S. Immunoglobulin G and immunoglobulin M enzyme-linked immunosorbent assays and defined toxoplasmosis serological patterns. Infect Immun. 1978 Jul;21(1):55–58. doi: 10.1128/iai.21.1.55-58.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dissous C., Grzych J. M., Capron A. Schistosoma mansoni surface antigen defined by a rat monoclonal IgG2a. J Immunol. 1982 Nov;129(5):2232–2234. [PubMed] [Google Scholar]
  4. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry. 1971 Sep;8(9):871–874. doi: 10.1016/0019-2791(71)90454-x. [DOI] [PubMed] [Google Scholar]
  5. Godson G. N., Ellis J., Svec P., Schlesinger D. H., Nussenzweig V. Identification and chemical synthesis of a tandemly repeated immunogenic region of Plasmodium knowlesi circumsporozoite protein. Nature. 1983 Sep 1;305(5929):29–33. doi: 10.1038/305029a0. [DOI] [PubMed] [Google Scholar]
  6. Handman E., Goding J. W., Remington J. S. Detection and characterization of membrane antigens of Toxoplasma gondii. J Immunol. 1980 Jun;124(6):2578–2583. [PubMed] [Google Scholar]
  7. Johnson A. M., McDonald P. J., Neoh S. H. Monoclonal antibodies to Toxoplasma cell membrane surface antigens protect mice from toxoplasmosis. J Protozool. 1983 May;30(2):351–356. doi: 10.1111/j.1550-7408.1983.tb02929.x. [DOI] [PubMed] [Google Scholar]
  8. Johnson A. M., McNamara P. J., Neoh S. H., McDonald P. J., Zola H. Hybridomas secreting monoclonal antibody to Toxoplasma gondii. Aust J Exp Biol Med Sci. 1981 Jun;59(Pt 3):303–306. doi: 10.1038/icb.1981.23. [DOI] [PubMed] [Google Scholar]
  9. Kasper L. H., Crabb J. H., Pfefferkorn E. R. Purification of a major membrane protein of Toxoplasma gondii by immunoabsorption with a monoclonal antibody. J Immunol. 1983 May;130(5):2407–2412. [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Naot Y., Remington J. S. An enzyme-linked immunosorbent assay for detection of IgM antibodies to Toxoplasma gondii: use for diagnosis of acute acquired toxoplasmosis. J Infect Dis. 1980 Nov;142(5):757–766. doi: 10.1093/infdis/142.5.757. [DOI] [PubMed] [Google Scholar]
  12. Rodriguez C., Afchain D., Capron A., Dissous C., Santoro F. Major surface protein of Toxoplasma gondii (p30) contains an immunodominant region with repetitive epitopes. Eur J Immunol. 1985 Jul;15(7):747–749. doi: 10.1002/eji.1830150721. [DOI] [PubMed] [Google Scholar]
  13. Sethi K. K., Endo T., Brandis H. Hybridomas secreting monoclonal antibody with specificity for Toxoplasma gondii. J Parasitol. 1980 Apr;66(2):192–196. [PubMed] [Google Scholar]
  14. Sharma S. D., Mullenax J., Araujo F. G., Erlich H. A., Remington J. S. Western Blot analysis of the antigens of Toxoplasma gondii recognized by human IgM and IgG antibodies. J Immunol. 1983 Aug;131(2):977–983. [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Zavala F., Cochrane A. H., Nardin E. H., Nussenzweig R. S., Nussenzweig V. Circumsporozoite proteins of malaria parasites contain a single immunodominant region with two or more identical epitopes. J Exp Med. 1983 Jun 1;157(6):1947–1957. doi: 10.1084/jem.157.6.1947. [DOI] [PMC free article] [PubMed] [Google Scholar]

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