Multiplex RT-PCR with broad-range primers and an exogenous internal amplification control for the detection of honeybee viruses in bumblebees.
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.
Meeus
Ivan
I
Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium. ivan.meeus@ugent.be
Smagghe
Guy
G
Siede
Reinhold
R
Jans
Kris
K
de Graaf
Dirk C
DC
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*
D012367:RNA, Viral; D012321:DNA-Directed RNA Polymerases