Hostname: page-component-cd9895bd7-fscjk Total loading time: 0 Render date: 2024-12-29T00:51:28.320Z Has data issue: false hasContentIssue false

A monoclonal-based IgM capture ELISA for detection of antibodies to 22 and 41 kDa membrane antigens of Toxoplasma gondii

Published online by Cambridge University Press:  06 April 2009

J. L. K. Wee
Affiliation:
Department of Microbiology, National University of Singapore, 10 Kent Ridge Road, Singapore0511
L. C. Ho
Affiliation:
Department of Microbiology, National University of Singapore, 10 Kent Ridge Road, Singapore0511
E. H. Yap
Affiliation:
Department of Microbiology, National University of Singapore, 10 Kent Ridge Road, Singapore0511
M. Singh
Affiliation:
Department of Microbiology, National University of Singapore, 10 Kent Ridge Road, Singapore0511

Summary

A murine monoclonal antibody which reacts to 22 and 41 kDa Toxoplasma gondii surface antigens was employed in an IgM capture enzyme-linked immunosorbent assay (ELISA). A total of 125 patients' sera were tested in the monoclonal-based assay. When compared with a commercial ELISA test (Abbott Toxo-M EIA) which uses polyclonal anti-T. gondii antibodies, good correlation (Pearsons coefficient r = 0.91) was observed. The specificity of the assay was studied by testing a panel of control sera obtained from healthy individuals and blood transfusion donors; all sera gave negative results. Serum samples positive for T. gondii antibodies were treated with 2-mercaptoethanol (2-ME) to demonstrate the specificity of the test for IgM antibodies. Reactivity of these sera was lost after the treatment. The test is not subject to interference by rheumatoid factor as sera positive for rheumatoid factor were negative in the assay. Reproducibility was good with the coefficients of variation for within-day tests below 10% and not exceeding 18% for day-to-day tests. The monoclonal-based assay is simple to perform and appears to be a viable test for diagnosis of T. gondii infection.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1992

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Anderson, L. J., Tsou, C., Parker, R. A., Chorba, T. L., Wulff, H., Tattersall, P. & Mortimer, P. P. (1986). Detection of antibodies and antigens of human parvovirus B19 by enzyme-linked immunosorbent assay. Journal of Clinical Microbiology 24, 522–6.CrossRefGoogle ScholarPubMed
Araujo, F. G., Barnett, E. V., Gentry, L. O. & Remington, J. S. (1971). False-positive anti-Toxoplasma fluorescent-antibody test in patients with antinuclear antibodies. Applied Microbiology 22, 270–5.CrossRefGoogle ScholarPubMed
Araujo, F. G., Dubey, J. P. & Remington, J. S. (1984). Antigenic similarity between coccidian parasite Toxoplasma gondii and Hammondia. Journal of Protozoology 31, 145–7.CrossRefGoogle ScholarPubMed
Balfour, A. H., Harford, J. P. & Goodall, M. (1987). Use of monoclonal antibodies in an ELISA to detect IgM class antibodies specific for Toxoplasma gondii. Journal of Clinical Pathology 40, 853–7.CrossRefGoogle Scholar
Camargo, M. E., Ferreira, A. W., Mineo, J. R., Takaguti, J. R. & Nakahara, O. S. (1978). Immunoglobulin G and immunoglobulin M enzyme-linked immunosorbent assays and defined toxoplasmosis serological patterns. Infection and Immunity 21, 55–8.CrossRefGoogle ScholarPubMed
Cesbron, J. Y., Capron, A., Ovalaque, G. & Santoro, F. (1985). Use of a monoclonal antibody in a double sandwich ELISA for the detection of IgM antibodies to Toxoplasma gondii major surface protein (P30). Journal of Immunological Methods 83, 151–8.CrossRefGoogle Scholar
Couvreur, G., Sadak, A., Fortier, B. & Dubremetz, J. F. (1988). Surface antigens of Toxoplasma gondii. Parasitology 97, 110.CrossRefGoogle ScholarPubMed
Galfre, G. & Milstein, C. (1981). Preparation of monoclonal antibodies: Strategies and procedures. Methods in Enzymology 73, 352.CrossRefGoogle ScholarPubMed
Gerna, G., Zannino, M., Revello, M. G., Petruzzelli, E. & Dovis, M. (1987). Development and evaluation of a capture enzyme-linked immunosorbent assay for determination of rubella immunoglobulin M using monoclonal antibodies. Journal of Clinical Microbiology 25, 1033–8.CrossRefGoogle ScholarPubMed
Handman, E., Goding, J. W. & Remington, J. S. (1980). Detection and characterisation of membrane antigens of Toxoplasma gondii. Journal of Immunology 124, 2578–83.CrossRefGoogle ScholarPubMed
Hughes, H. P. A. (1985). Toxoplasmosis: The need for improved techniques and accurate risks assessment. Current Topics in Microbiology and Immunology 120, 106–9.Google Scholar
Huskinson, J., Stepick-Biek, P. N., Araujo, F. G., Thulliez, P., Suzuki, Y. & Remington, J. S. (1989). Toxoplasma antigens recognized by immunoglobulin G subclasses during acute and chronic infection. Journal of Clinical Microbiology 27, 2031–8.CrossRefGoogle ScholarPubMed
Hyde, B., Barnett, E. B. & Remington, J. S. (1975). Method for differentiation of nonspecific from specific Toxoplasma IgM fluorescent antibodies in patients with rheumatoid factor. Proceedings of the Society of Experimental Biology and Medicine 148, 1184–8.CrossRefGoogle ScholarPubMed
Kasper, L. H., Crabb, J. H. & Pffefferkorn, E. R. (1983). Purification of a major membrane protein of Toxoplasma gondii by immunoabsorption with a monoclonal antibody. Journal of Immunology 130, 2407–12.CrossRefGoogle Scholar
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Lunde, M. N. & Jacobs, L. (1965). Antigen relationship of Toxoplasma gondii and Besnoitia jellisoni. Journal of Parasitology 51, 273–6.CrossRefGoogle ScholarPubMed
Luger, A. F. H. (1988). Serological diagnosis of syphilis. In Immunological Diagnosis of Sexually Transmitted Diseases (ed. Young, H. & McMillan, A.), pp. 249274. New York: Marcel Dekker Inc.Google Scholar
Lyng, J. (1987). An international standard for anti-Toxoplasma IgM antibodies. In Technical Report Series No. 745, New WHO Publication, pp. 122. Geneva: World Health Organization.Google Scholar
Naot, Y. & Remington, J. S. (1980). An enzyme-linked immunosorbent assay for detection of IgM antibodies to Toxoplasma gondii: use for the diagnosis of acute acquired toxoplasmosis. Journal of Infectious Diseases 142, 757–66.CrossRefGoogle ScholarPubMed
Partanen, P., Turunen, H. J., Paasivuo, R. T. A. & Leinikki, P. O. (1974). Immunoblot analysis of Toxoplasma gondii antigens by human immunoglobulins G, M and A antibodies at different stages of infection. Journal of Clinical Microbiology 20, 133–5.CrossRefGoogle Scholar
Pedersen, N. S., Petersen, C. S. & Axelsen, N. H. (1982). Enzyme-linked immunosorbent assay for detection of immunoglobulin M antibody against the Reiter treponeme flagellum in syphilis. Journal of Clinical Microbiology 16, 608–14.CrossRefGoogle ScholarPubMed
Potasman, I., Araujo, F. G., Desmont, G. & Remington, J. S. (1986 a). Analysis of Toxoplasma gondii antigens recognized by human sera obtained before and after acute infection. Journal of Infectious Diseases 154, 650–7.CrossRefGoogle ScholarPubMed
Potasman, I., Araujo, F. G. & Remington, J. S. (1986 b). Toxoplasma antigens recognized by naturally occurring human antibodies. Journal of Clinical Microbiology 24, 1050–4.CrossRefGoogle ScholarPubMed
Santoro, F., Charif, H. & Capron, A. (1986). The immunodominant epitope of the major membrane tachyzoite protein (P30) of Toxoplasma gondii. Parasite Immunology 8, 631–9.CrossRefGoogle ScholarPubMed
Sharma, S. D., Mullenax, J., Araujo, F. G., Erlich, H. A. & Remington, J. S. (1983). Western blot analysis of the antigens of Toxoplasma gondii recognized by human IgM and IgG antibodies. Journal of Immunology 131, 977–83.CrossRefGoogle ScholarPubMed
Sibley, L. D., Krahenbuhl, J. L., Adams, G. M. W. & Weidner, E. (1986). Toxoplasma modifies macrophage phagosomes by secretion of a vesicular network rich in surface proteins. Journal of Cell Biology 103, 867–74.CrossRefGoogle Scholar
Towbin, H., Staehlin, T. & Gordon, J. (1978). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, USA 76, 4350–4.CrossRefGoogle Scholar
Van Loon, A. M., Van Der Logt, J. T. M., Heessen, F. W. A. & Van Der Veen, J. (1983). Enzyme-linked immunosorbent assay that uses labeled antigen for the detection of immunoglobulin M and A antibodies in toxoplasmosis: comparison with indirect immunofluorescence and double-sandwich enzyme-linked immunosorbent assay. Journal of Clinical Microbiology 17, 9971004.CrossRefGoogle Scholar
Weiss, L. M., Udem, S. A., Tanowitz, H. & Wittner, M. (1988). Western blot analysis of the antibody response of patients with AIDS and Toxoplasma encephalitis: Antigenic diversity among Toxoplasma strains. Journal of Infectious Diseases 157, 713.CrossRefGoogle ScholarPubMed