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Characteristics of Toxoplasma-gondii-tachyzoites from different culture systems

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Harald Klein1, Martina Anduleit1, Monika Bornhak1, Matthias Fischer1, Uwe Gross2, Bettina Löschner1, Sven Nicol1, Ingrid Reiter-Orwona3, Nadja Zyto1 und Thomas Montag-Lessing1
1Paul-Ehrlich-Institut, D-Langen, 2Institut für Hygiene und Mikrobiologie der Universität, D-Würzburg, 3Institut für Medizinische Parasitologie der Universität D-Bonn
Toxoplasma gondii is an intracellular protozoan parasite of worldwide distribution. Parasites that habour a complete antigenic profile, that is necessary for the serological diagnosis of human Toxoplasma infections, are provided by in vivo culture methods only. It seems that the host immune pressure is responsible for the expression of a total antigen pattern. Thus, in most laboratories the asexual proliferative stage of the parasite, the tachyzoite, is maintained by successive intraperitoneal passages in highly susceptible animals such as mice.
We would like to develop an in vitro method to provide sufficient amounts of high quality parasite antigen suitable for diagnosis of Toxoplasmosis. Using the RAPD-PCR (random amplified polymorphic DNA-PCR) technique, we were able to show that during different culture conditions (in vivo and in vitro culture) the parasites undergo no clonal selection. That means the entire parasite population changes the protein expression pattern due to the in vitro culture conditions. Based on the result described previously the work could be continued as follows. One strategy could be to simulate the host immune pressure during the in vitro cultivation of the tachyzoites. A more convinced approach may be the production of recombinant parasite antigens useful for diagnosis of a Toxoplasma infection in human adults and newborns.

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