Submitted to: Journal of Virological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 11, 2009
Publication Date: December 11, 2009
Citation: Boncristiani Jr, H.F., Diprisco, G., Pettis, J.S., Hamilton, M.C., Chen, Y. 2009. Molecular approaches to the Analysis of Virus Replication and Pathogenesis in Honey Bees, Apis mellifera. Journal of Virological Methods. 6:221. Interpretive Summary: Viruses pose a serious threat to the health and well-being of honey bees, the most economically valuable pollinators of the agricultural system. Recently, honey bee viruses have drawn significant attention to scientific community and beekeeping industry due to the serious diseases that viruses cause in honey bees. The precise identification of the virus replication is an important step toward the understanding of viral disease mechanisms in honey bees. We conducted a study to develop a novel approach that allows for reliable and rapid identification of the virus replication in honey bees. This study should be a significant contribution to the design of appropriate control measures for bee populations at risk of virus infections. The results have relevance to scientists, regulators, and beekeepers seeking to mitigate recent bee losses.
Technical Abstract: For years, understanding of the pathogenetic mechanisms that underlie honey bee viral diseases has been severely hindered because of the lack of a cell culture system for virus propagation. As a result, it is very imperative to develop new methods that would permit the in vitro pathogenesis study of honey bee viruses. The identification of virus replication is an important step towards the understanding of the pathogenecity of viruses in their respective hosts. We developed a novel strand specific PCR-based method for analysis of honey bee Deformed Wing Virus (DWV) replication. The results show that the method developed in our study allows reliable identification of the virus replication and solves the problems of falsely-primed cDNAs amplification that commonly exist in the current system. Using TaqMan real time quantitative PCR incorporated with biotinylated primers and magnetic beads purification step, we characterized the replication and tissue tropism of DWV infection in honey bees and parasitic mites, Varroa. This study forms a model system for studying bee virus replication, pathogenesis and immunity. The strategy reported in the present study should contribute to the better understanding of virus pathogenecity in honey bees and to the design of appropriate disease control strategies.