An immunochemical method for quantitation of Epinotia aporema granulovirus (EpapGV).

ABSTRACT

Epinotia aporema granulovirus (EpapGV) is a baculovirus that affects E. aporema larvae and has proven to be a good candidate for the biocontrol of this important pest in South America. As part of the quality control of the production of a bioinsecticide based on EpapGV, a sensitive method was developed for the detection and quantitation of the virus. To this end, we used the major occlusion body (OB) protein (granulin) to generate polyclonal antibodies in rabbits. Purified IgG fractions from hyperimmune sera were labeled with biotin and used as detecting antibodies in a double antibody sandwich enzyme linked immunosorbent assays (ELISA). No cross-reactivity was detected with any of the nucleopolyhedroviruses (NPV) tested in this study, while a minor degree of reactivity was observed with the closely related Cydia pomonella granulovirus (CpGV). The performance of the ELISA was satisfactory in terms of sensitivity, detecting as little as 0.53 ng/ml of EpapGV granulin in suspensions of purified virus OB. This represented 2.0x10(4) OB/ml. Granulin was also detected in complex and highly diluted bioinsecticidal formulate mixtures. In time course experiments, the virus was detected as early as 24 h post infection (p.i.). The results of the studies demonstrate that this method is a convenient, rapid and inexpensive alternative for routine detection and quantitation of EpapGV.