BeeDoctor, a versatile MLPA-based diagnostic tool for screening bee viruses

• Published in: PloS one 2012-10, Vol.7 (10), p.e47953-e47953
• Main Authors: De Smet, Lina, Ravoet, Jorgen, de Miranda, Joachim R, Wenseleers, Tom, Mueller, Matthias Y, Moritz, Robin F A, de Graaf, Dirk C
• Format: Article
• Language: English
• Subjects: Agriculture; Animals; Annan veterinärmedicin; Apis mellifera; Assaying; Bees; Bees - virology; Biofilms; Biology; Colonies; Deformation; Diagnostic software; Diagnostic systems; Ecology; Ekologi; Epidemiology; European honeybee; High-throughput screening; Honey; Infections; Insect viruses; Insect Viruses - classification; Insect Viruses - genetics; Laboratories; Medical diagnosis; Medical screening; Multiplex Polymerase Chain Reaction - methods; Multiplexing; Mycobacterium tuberculosis; Other Veterinary Science; Pathogens; Penicillium; Physiology; Polymerase chain reaction; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction - methods; Ribonucleic acid; RNA; RNA viruses; RNA Viruses - classification; RNA Viruses - genetics; RNA, Viral - genetics; Screening; Tuberculosis; Varroa destructor; Veterinary Science; Viral infections; Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0047953

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 bee viruses or the negative strand, which is indicative for active viral replication. Due to its sensitivity and specificity, the MLPA assay is a useful tool for rapid diagnosis, pathogen characterization, and epidemiology of viruses in honeybee populations. "BeeDoctor" was used for screening 363 samples from apiaries located throughout Flanders; the northern half of Belgium. Using the "BeeDoctor", virus infections were detected in almost eighty percent of the colonies, with deformed wing virus by far the most frequently detected virus and multiple virus infections were found in 26 percent of the colonies.