Rapid Baculovirus Detection, Identification, and Serological Classification by Western Blotting-ELISA using a Monoclonal Antibody

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Abstract
A hybridoma cell line, called 3D10, was established which secretes monoclonal antibodies against the 42K protein of Autographa californica nuclear polyhedrosis virus (Ac NPV). Following separation of a complex protein mixture by denaturating sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretic protein transfer to a nitrocellulose filter, this monoclonal antibody is still able to react with its antigen. The antigen-antibody complex can be stained by indirect enzyme-linked immunosorbent assay (ELISA) using horseradish peroxidase-conjugated rabbit anti-mouse serum. The Western blot-ELISA described is extremely sensitive, being able to detect as little as 10 ng of Ac NPV or 3 .times. 105 polyhedral inclusion bodies. Using this method it is possible to find traces of Ac NPV in very crude materials, e.g., impure polyhedra, infectious hemolymph and larval homogenates. This monoclonal antibody is very specific and reacts only with isolates of Ac NPV and Galleria mellonella NPV. No reactions were found against 11 other NPV isolates. The implications of these findings for baculovirus identification and classification are discussed.