Preparation of the Antibody Against EupsNPV Polyhedrin and Its Utilization in Detection of the Viral Pesticide

The 741 bp coding region of polyhedrin gene (ph) was amplified from Euproctis pseudoconspersa nucleopolyhedrovirus(EupsNPV)genome and its prokaryotic expression vector pET-28-a-Ep-ph was constructed. Then the prokaryotic expression plasmid was transformed into Escherichia.coli. BL21(DE3)for expression. The expressed fusion protein was separated by SDS-PAGE and retrieved from the gel. New Zealand white rabbit was immunized with the purified fusion protein and the antiserum against ph with a titre of 6.4×104 was prepared. Western blotting indicated that the prepared polyclonal antibody was specific for polyhedrin. The EupsNPV was detected by indirect ELISA and its regression equation was obtained: y=0.4152x-0.8299 with correlation coeffient r=0.9897(P0.01). Indirect ELISA detection showed that the antibody could be used for quantitative detection of the NPV in a commercial bio-pesticide and provided an accurate and effective method for detection.