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The rhoptry kinase (ROPK) and the rhoptry protein 3 (ROP3) are very important components of the parasitophorous vacuolar membrane and the rhoptry contents, respectively, of the tachyzoite stage of Toxoplasma gondii. In this work, the recombinant T. gondii ROPK and ROP3 proteins were expressed in Escherichia coli, and used as antigens to study the generation of antibody responses in rabbits and humans experimentally infected with T. gondii. The enzyme-linked immunosorbent assay (ELISA) was used to detect the antibodies to the ROPK and ROP3 in the sera of rabbits and humans. The total IgG antibodies were detected by indirect ELISA. The total IgG and IgM antibodies were also detected by direct ELISA using immobilized antigens. The results indicated that there was a significant difference between the absorbance values at 405 nm of all rabbit sera samples in the indirect ELISA using the ROPK and ROP3 proteins and those of positive controls. The positive rate of ROPK and ROP3 indirect ELISA for acute and chronic toxoplasmosis was higher than that of the indirect ELISA using whole tachyzoites.
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