beedoctor, a versatile mlpa-based diagnostic tool for screening bee virusesbeedoctor,多功能mlpa-based蜜蜂病毒筛查的诊断工具.pdfVIP

BeeDoctor, a Versatile MLPA-Based Diagnostic Tool for Screening Bee Viruses 1 1 2 3 4 Lina De Smet *, Jorgen Ravoet , Joachim R. de Miranda , Tom Wenseleers , Matthias Y. Mueller , Robin F. A. Moritz4, Dirk C. de Graaf1 1 Laboratory of Zoophysiology, Department of Physiology, Ghent University, Ghent, Belgium, 2 Department of Ecology, Swedish University of Agricultural Sciences, ¨ ¨ ¨ Uppsala, Sweden, 3 Laboratory of Socioecology and Social Evolution, KU Leuven, Leuven, Belgium, 4 Institut fur Biologie, Martin Luther Universitat Halle-Wittenberg, Halle/Saale, Germany Abstract The long-term decline of managed honeybee hives in the world has drawn significant attention to the scientific community and bee-keeping industry. A high pathogen load is believed to play a crucial role in this phenomenon, with the bee viruses being key players. Most of the currently characterized honeybee viruses (around twenty) are positive stranded RNA viruses. Techniques based on RNA signatures are widely used to determine the viral load in honeybee colonies. High throughput screening for viral loads necessitates the development of a multiplex polymerase chain reaction approach in which different viruses can be targeted simultaneously. A new multiparameter assay, called ‘‘BeeDoctor’’, was developed based on multiplex-ligation probe dependent amplification (MLPA) technology. This assay detects 10 honeybee viruses in one reaction. ‘‘BeeDoctor’’ is also able to screen selectively for either the positive strand of the targeted RNA