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Title:   Author  item
                                            Chen, Yanping

                                                         - *Judy*item
                                            Zhao, Yanitem
                                            Hammond, Johnitem
                                            Hsu, Hei Tiitem
                                            Evans, Jayitem
                                            Feldlaufer, Mark
  Submitted to: Journal of Invertebrate Pathology
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 7/19/2004
                                    Publication  Date: 7/19/2004
                                    Citation: Chen, Y.P., Zhao, Y., Hammond, J., Hsu, H., Evans, J.D., Feldlaufer, M.F. 2004. Multiple virus infections in the honey bee and genome divergence of honey bee viruses. Journal of Invertebrate Pathology.87:84-93.Interpretive Summary: Honey bees are important beneficial insects that can be infected by numerous disease agents, including viruses. Honey bees can often suffer from multiple virus infections and it is difficult to diagnose these infections in the field since many of the viruses produce no apparent symptoms. We have developed a molecular method that allows the simultaneous detection of multiple bee viruses in a single reaction.   This method is quick, accurate and cost-effective. It can be used by apiary inspectors to monitor honey bee colonies for viruses and isolate and quarantine infected colonies to prevent the spread of disease. It can also be used by regulatory agencies to determine the virus status of bee colonies from countries interested in exporting bees to the United States.
                                Technical Abstract: Using uniplex RT-PCR we screened honey bee colonies for the presence of several bee viruses, including Black Queen Cell virus (BQCV), Deformed Wing virus (DWV), Kashmir Bee virus (KBV), and Sacbrood virus (SBV), and described the detection of mixed virus infections in bees from these colonies.  We report for the first time that individual bees can harbor four viruses simultaneously. We also developed a multiplex RT-PCR assay for the simultaneous detection of multiple bee viruses. The feasibility and specificity of the multiplex RT-PCR assay suggests that this assay is an effective tool for simultaneous examination of mixed virus infections in bee colonies and would be useful for the diagnosis and surveillance of honey bee viral diseases in the field and laboratory.  Phylogenetic analysis of putative helicase and RNA-dependent RNA polymerase (RdRp) encoded by viruses reveal that DWV and SBV fall into a distinct group, whereas KBV and BQCV belong to a distinct lineage with other picorna-like viruses that infect plants, insects and vertebrates. Results from field surveys of these viruses indicate that mixed infections of BQCV, DWV, KBV, and SBV in the honey bee probably arise due to broad geographic distribution of viruses.


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