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Toxoplasmosis Diagnostics

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Development of the Technology of the Enzyme Immunoassay Kits for Toxoplasmosis Diagnostics with Use of Purified of Toxoplasma Gondii Proteins

Tech Area / Field

  • BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
  • BIO-CHM/Biochemistry/Biotechnology
  • BIO-MIB/Microbiology/Biotechnology

Status
8 Project completed

Registration date
08.07.1996

Completion date
08.02.2002

Senior Project Manager
Pobedimskaya D D

Leading Institute
State Research Center of Virology and Biotechnology VECTOR, Russia, Novosibirsk reg., Koltsovo

Collaborators

  • US Department of Agriculture, USA, MD, Beltsville

Project summary

The goal of the present project is to develop a set of kits for evident diagnostics of acute phase of toxoplasmosis using enzyme immunoassay. Toxoplasmosis is a widespread disease caused by an intracellular protozoan parasite, Toxoplasma gondii. It is generally harmless for adults with exception to the cases of immunodeficiency, especially AIDS and bonemarrow or heart-transplanted patients. In healthy population toxoplasmosis is mostly dangerous for congenitally infected children. Acquired acute toxoplasmosis in the pregnant women may result in spontaneous aborts or very serious diseases for their offspring. Therefore, it is of utmost importance to detect acute acquired infection in women during pregnancy and allow timely treatment to prevent infection of the fetus.

Clinical symptoms of acute toxoplasmosis are, as a rule, very minor and indefinite, and reliable diagnosis may be obtained by serological examination. At present ELISA is the mostly widespread method because of its sensitivity, reproducibility and simplicity. A number of ELISA kits are commercially available.

Unfortunately, these test-systems are often unable to give unequivocal Conclusions about the date of infection. The acute infection is classically characterized by the presence of toxoplasma-specific IgM antibodies. However, in some cases IgM antibodies were found to persist in sera for long time after the onset of the infection. In other cases only IgG antibodies may be produced in sera at acute phase. Finally, 'false positive' sera were found to contain rheumatoid factors, or antibodies cross-reacting with other microorganisms. The present project is planned to realize this way.

ELISA kits based on the inpidual proteins are widely used in diagnostics of different virus and bacterial diseases. This kit was shown to be more sensitive and specific. In the case of parasitic diseases, for example toxoplasmosis, the creation of such kits is in the beginning stage, because toxoplasma tachizoits is the more complex objects, containing much different protein with unknown functions. Scientists, as a rule, have for theirs investigation taxizoits from the mouse ascites. This preparates contain impurities of the host proteins, because it is difficult to get unequivocal data.

In our institute it was created effective methods of toxoplazma cultivation, which have been allowed to prepare of taxizoites with 90 % purity. By this method it is possible to get need to ELISA kits production quantity of toxoplasma antigens.

Expected result of project is as follows:


- to develop the purification technique for immunologically important toxoplasma antigens;
- to construct immunoenzyme test-system for detection of IgG and IgM antibodies against toxoplasma antigens;
- to construct immunoblotting kit for analysis of toxoplasma-specific antibodies;
- to prepared scientific and engineering specification.

Participation of foreign collaborators may conclude in the trials of ELISA kits and in its marketing.


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