A hybridoma cell line, called 3D10, has been established which secretes monoclonal antibodies against the 42K protein of nuclear polyhedrosis virus (Ac NPV). Following separation of a complex protein mixture by denaturating SDS-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 × 10 polyhedral inclusion bodies. Using this method it is possible to find traces of Ac NPV in very crude materials, e.g. impure polyhedra, infectious haemolymph, and larval homogenates. On the other hand, this monoclonal antibody is very specific and reacts only with isolates of Ac NPV and NPV. No reactions were found against eleven other NPV isolates. The implications of these findings for baculovirus identification and classification are discussed.


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