Production and application of monoclonal antibodies against duck plague virus

Abstract

Monoclonal antibodies (Mabs) were raised against the vaccine strain of DPV and three strains of DPV Viz, Vaccine (DPV-V), IVRI (DPV-I) and Alleppy strain (DPV-A) were used to raise polyclonal serum in the present investigation. DPV-V was revived in 11 day old chicken embryo and the embryo death was recorded Tour to five days PI- with congestion all over the body and spleen and necrotic foci in liver. The cytopalhy in CEF cell culture observed was rounding and clumping of the cells, syncylium formation and bridge formation with extensive vacuolation in the Cytoplasm. The detachment of the cells was observed at 120 h PI. DPV-I a virulent strain was inoculated in the ducklings, death was recorded in all the inoculated birds with extensive hemorrhages on serous membranes, muscles and visceral organs. Necrotic foci on liver,' enlargement and congestion of liver, and spleen, and white hecrotic foci in the gizzard were evident. The virus was further passaged in DDE and cultivated in bulk in DEF cell culture. The DPV-V and DPV-A were titrated in CEF cell culture and the TC1D J0 was 4.7 X 105 per ml of the inoculum Tor DPV-V and 3.2 X 10"1 for DPV-A. DPV-I cultivated in DEF cell culture had a TCID50 of 1X10 3'per ml of the inoculum All (lie strains were partially purified at 100000 g for 4.5 h at 4" C in Beckman ultra centrifuge and the protein concentration oF the virus was estimated by biuret method and was found to be 11 mg, 8 mg and 7 mg for DPV-V, A and 1 respectively. All the three strains of DPV were inoculated in mice to raise polyclonal serum. Four mice out of five inoculated with DPV-V showed ELISA litres more than 1:12800, one mouse showed a titre of 1:6400. The mice inoculated with DPV-A showed a titre of more than 1,12800 and those inoculated with DPV-I, 1:6400 ELISA ’Was1’ used to test the sera samples of the mice inoculated with DPV strains. The test was found to be highly sensitive, easy to perform and less time consuming. The test therefore can be recommended for routine diagnosis of DPV