Development of Diagnostic Systems on the Piroplasma lnfection in Cattle and Canine

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1. Preparations of Monoclonal antibodies for protozoan proteins: Mab for protozoa of Babesia spp. And Theileria spp., infected within erythrocytes of cattle and canine were prepared. In a many of Mab for Babesia (B.) gibsoni, there were isolated two Mab recognized proteins of 30 kDa and 24.5 kDa in the protozoa. A Mab for B. ovata recognized proteins of 19 kDa and two Mab for Theileria (T.) sergenti recognized proteins of 23 kDa and 32 kDa.2. Diagnostic methods using Mab for B. gibsoni(1) Development of enzyme-linked immunosorbent assay (ELISA) system; ELISA systems by sandwiche method were investigated using each Mab for proteins of 30 and 24.5 kDa in B. gibsoni. In their assay systems, when Mab for 30 kDa protein was used as a antibody of solid phase and Mab for 24.5 kDa protein was labeled with biotin and streptolysin, the sensitivity and specificity of the reaction showed excellent results. There was highly significant correlated between babesial soluble protein in serum of canine … More infected with B. gibsoni and percentages of protozoa in erythrocytes (r=0.879). And this ELISA was positive at a percentage above 0.05% parasitemia of protozoa.(2) Latex agglutination test; Using latex beads sensitized with Mab, the agglutination test using latex sensitized with Mab for 24.5 kDa protein showed most sensitive and specific reaction. The correlation with parasitemia was high significantly (r=0.832). But, this ELISA was negative at a percentage under 0.1% parasitemia of protozoa.3. Diagnostic methods using Mab for T. sergenti(1) Development of enzyme-linked immunosorbent assay (ELISA) system; Sandwiche ELISA systems using Mab for 30 and 24.5 kDa in T. sergenti also showed good reactions. In their assay systems, good reactional condition was observed by using Mab for 23 kDa protein.(2) Latex agglutination test; Using latex beads sensitized with Mab for 23 kDa protein, the agglutination test showed most sensitive and specific reaction. The correlation with parasitemia was high significantly (r=0.801). But, this ELISA was negative at a percentage under 0.2% parasitemia of protozoa. Less