Development and Preliminary Application of a Blocking ELISA for Detection of Porcine Toxoplasmosis
In this study,a blocking ELISA method for swine toxoplasmosis was developed based on the monoclonal antibody GRA7.Mice were immunized with the recombinant protein rGRA7 and then their spleen cells were taken for cell fusion.The resulting hybridomas were screened and 4 positive cell lines were found to stably produce monoclonal antibodies.The monoclonal antibodies against GRA7 protein was confirmed in IFA and Western blotting and used to develop a blocking ELISA method.The blocking ELISA was optimized for the concentration of the coating antigen and tested for its repeatability,cross-compatibility and conformity.The results showed that this blocking ELISA had good repeatability and high compliance with MAT.Subsequently,this method was used to test 288 clinical samples and the positive rate was 11.46％with 84.9％coincidence with the MAT.Collectively,a monoclonal antibody against GRA7 protein of Toxoplasma gondii was prepared for development of a blocking ELISA method,which might be used for the detection of swine toxoplasmosis.
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