Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1992 Jun;30(6):1436–1441. doi: 10.1128/jcm.30.6.1436-1441.1992

Improved serological diagnosis of Toxoplasma gondii infection by detection of immunoglobulin A (IgA) and IgM antibodies against P30 by using the immunoblot technique.

U Gross 1, T Roos 1, D Appoldt 1, J Heesemann 1
PMCID: PMC265306  PMID: 1624560

Abstract

Immunoglobulin M (IgM) and IgA antibodies against the major surface protein of Toxoplasma gondii were determined in a total of 195 human sera and five human cerebrospinal fluids by using a P30 membrane extract and the immunoblot technique. By using two different T. gondii strains (RH and BK) simultaneously as antigens, we were able to demonstrate diagnostically important strain-specific human antibody responses in 4.5% of the samples tested. A comparison of the immunoblot technique with an IgM immunocapture enzyme-linked immunosorbent assay demonstrated that the IgM and IgA immunoblot seems to be of advantage in the diagnosis of acute toxoplasmosis in certain groups of patients, especially in the diagnosis of cerebral toxoplasmosis in patients with AIDS. The immunoblot technique described is easy to perform and might be useful as an additional serological assay for routine diagnosis of T. gondii infections.

Full text

PDF
1440

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Araujo F. G., Remington J. S. IgG antibody suppression of the IgM antibody response to Toxoplasma gondii in newborn rabbits. J Immunol. 1975 Aug;115(2):335–338. [PubMed] [Google Scholar]
  2. Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
  3. Brandtzaeg P., Halstensen T. S., Kett K., Krajci P., Kvale D., Rognum T. O., Scott H., Sollid L. M. Immunobiology and immunopathology of human gut mucosa: humoral immunity and intraepithelial lymphocytes. Gastroenterology. 1989 Dec;97(6):1562–1584. doi: 10.1016/0016-5085(89)90406-x. [DOI] [PubMed] [Google Scholar]
  4. Burg J. L., Perelman D., Kasper L. H., Ware P. L., Boothroyd J. C. Molecular analysis of the gene encoding the major surface antigen of Toxoplasma gondii. J Immunol. 1988 Nov 15;141(10):3584–3591. [PubMed] [Google Scholar]
  5. Cesbron J. Y., Capron A., Ovlaque G., Santoro F. Use of a monoclonal antibody in a double-sandwich ELISA for detection of IgM antibodies to Toxoplasma gondii major surface protein (P30). J Immunol Methods. 1985 Oct 24;83(1):151–158. doi: 10.1016/0022-1759(85)90068-7. [DOI] [PubMed] [Google Scholar]
  6. Decoster A., Darcy F., Caron A., Capron A. IgA antibodies against P30 as markers of congenital and acute toxoplasmosis. Lancet. 1988 Nov 12;2(8620):1104–1107. doi: 10.1016/s0140-6736(88)90523-5. [DOI] [PubMed] [Google Scholar]
  7. Gross U., Müller W. A., Knapp S., Heesemann J. Identification of a virulence-associated antigen of Toxoplasma gondii by use of a mouse monoclonal antibody. Infect Immun. 1991 Dec;59(12):4511–4516. doi: 10.1128/iai.59.12.4511-4516.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gross U., Roggenkamp A., Janitschke K., Heesemann J. Improved sensitivity of the polymerase chain reaction for detection of Toxoplasma gondii in biological and human clinical specimens. Eur J Clin Microbiol Infect Dis. 1992 Jan;11(1):33–39. doi: 10.1007/BF01971268. [DOI] [PubMed] [Google Scholar]
  9. Grover C. M., Thulliez P., Remington J. S., Boothroyd J. C. Rapid prenatal diagnosis of congenital Toxoplasma infection by using polymerase chain reaction and amniotic fluid. J Clin Microbiol. 1990 Oct;28(10):2297–2301. doi: 10.1128/jcm.28.10.2297-2301.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Holliman R. E. Clinical and diagnostic findings in 20 patients with toxoplasmosis and the acquired immune deficiency syndrome. J Med Microbiol. 1991 Jul;35(1):1–4. doi: 10.1099/00222615-35-1-1. [DOI] [PubMed] [Google Scholar]
  11. Huskinson J., Stepick-Biek P. N., Araujo F. G., Thulliez P., Suzuki Y., Remington J. S. Toxoplasma antigens recognized by immunoglobulin G subclasses during acute and chronic infection. J Clin Microbiol. 1989 Sep;27(9):2031–2038. doi: 10.1128/jcm.27.9.2031-2038.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Huskinson J., Thulliez P., Remington J. S. Toxoplasma antigens recognized by human immunoglobulin A antibodies. J Clin Microbiol. 1990 Dec;28(12):2632–2636. doi: 10.1128/jcm.28.12.2632-2636.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Luft B. J., Remington J. S. AIDS commentary. Toxoplasmic encephalitis. J Infect Dis. 1988 Jan;157(1):1–6. doi: 10.1093/infdis/157.1.1. [DOI] [PubMed] [Google Scholar]
  15. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  16. Naot Y., Desmonts G., Remington J. S. IgM enzyme-linked immunosorbent assay test for the diagnosis of congenital Toxoplasma infection. J Pediatr. 1981 Jan;98(1):32–36. doi: 10.1016/s0022-3476(81)80528-8. [DOI] [PubMed] [Google Scholar]
  17. Partanen P., Turunen H. J., Paasivuo R. T., Leinikki P. O. Immunoblot analysis of Toxoplasma gondii antigens by human immunoglobulins G, M, and A antibodies at different stages of infection. J Clin Microbiol. 1984 Jul;20(1):133–135. doi: 10.1128/jcm.20.1.133-135.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Potasman I., Araujo F. G., Remington J. S. Toxoplasma antigens recognized by naturally occurring human antibodies. J Clin Microbiol. 1986 Dec;24(6):1050–1054. doi: 10.1128/jcm.24.6.1050-1054.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Santoro F., Afchain D., Pierce R., Cesbron J. Y., Ovlaque G., Capron A. Serodiagnosis of toxoplasma infection using a purified parasite protein (P30). Clin Exp Immunol. 1985 Nov;62(2):262–269. [PMC free article] [PubMed] [Google Scholar]
  21. Santoro F., Charif H., Capron A. The immunodominant epitope of the major membrane tachyzoite protein (P30) of Toxoplasma gondii. Parasite Immunol. 1986 Nov;8(6):631–639. doi: 10.1111/j.1365-3024.1986.tb00876.x. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Stepick-Biek P., Thulliez P., Araujo F. G., Remington J. S. IgA antibodies for diagnosis of acute congenital and acquired toxoplasmosis. J Infect Dis. 1990 Jul;162(1):270–273. doi: 10.1093/infdis/162.1.270. [DOI] [PubMed] [Google Scholar]
  24. Verhofstede C., Van Renterghem L., Plum J. Evaluation of immunoblotting for the detection of Toxoplasma gondii immunoglobulin M antibodies. Eur J Clin Microbiol Infect Dis. 1990 Nov;9(11):835–837. doi: 10.1007/BF01967387. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Wanke C., Tuazon C. U., Kovacs A., Dina T., Davis D. O., Barton N., Katz D., Lunde M., Levy C., Conley F. K. Toxoplasma encephalitis in patients with acquired immune deficiency syndrome: diagnosis and response to therapy. Am J Trop Med Hyg. 1987 May;36(3):509–516. doi: 10.4269/ajtmh.1987.36.509. [DOI] [PubMed] [Google Scholar]
  27. Weiss L. M., Udem S. A., Tanowitz H., Wittner M. Western blot analysis of the antibody response of patients with AIDS and toxoplasma encephalitis: antigenic diversity among Toxoplasma strains. J Infect Dis. 1988 Jan;157(1):7–13. doi: 10.1093/infdis/157.1.7. [DOI] [PubMed] [Google Scholar]
  28. Werner H. Der Nachweis von Toxoplasma gondii durch den Tierversuch. II. Methodik, Erfahrungen und Ergebnisse mit den im Robert Koch-Institut angewandten Anzüchtungsverfahren. Z Tropenmed Parasitol. 1966 Oct;17(3):328–336. [PubMed] [Google Scholar]
  29. Wong B., Gold J. W., Brown A. E., Lange M., Fried R., Grieco M., Mildvan D., Giron J., Tapper M. L., Lerner C. W. Central-nervous-system toxoplasmosis in homosexual men and parenteral drug abusers. Ann Intern Med. 1984 Jan;100(1):36–42. doi: 10.7326/0003-4819-100-1-36. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES