title: Multiplex RT-PCR with broad-range primers and an exogenous internal amplification control for the detection of honeybee viruses in bumblebees.
abstract: Bumblebees are commercially reared and transported worldwide mainly for pollination of greenhouse tomatoes. Three honeybee viruses have been reported in bumblebees: Acute bee paralysis virus, Kashmir bee virus and Deformed wing virus. We developed a multiplex RT-PCR with primers designed on highly conserved regions of the RNA-dependent RNA polymerase in order to detect a maximum range of viral variants. Rearing facilities and governmental organizations can now thoroughly screen bumblebee colonies with a cost-effective technique with an integrated internal amplification control (IAC) implementable in laboratories that strive for International Organization for Standardization (ISO) certification.
authors.affiliation: Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium. ivan.meeus@ugent.be
authors.forename: Ivan
authors.initials: I
authors.lastname: Meeus
authors.forename: Guy
authors.initials: G
authors.lastname: Smagghe
authors.forename: Reinhold
authors.initials: R
authors.lastname: Siede
authors.forename: Kris
authors.initials: K
authors.lastname: Jans
authors.forename: Dirk C
authors.initials: DC
authors.lastname: de Graaf
mesh_terms: D000818:Animals; D001516:Bees / Q000821:virology*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D056629:Dicistroviridae / Q000235:genetics*; D007304:Insect Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D010849:Picornaviridae / Q000235:genetics; D010850:Picornaviridae Infections / Q000175:diagnosis / Q000235:genetics / Q000662:veterinary*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D014777:Virus Diseases / Q000175:diagnosis / Q000235:genetics / Q000662:veterinary*
chemical_list: D012367:RNA, Viral; D012321:DNA-Directed RNA Polymerases