Multiplex RT-PCR with broad-range primers and an exogenous internal amplification control for the detection of honeybee viruses in bumblebees

J Invertebr Pathol. 2010 Oct;105(2):200-3. doi: 10.1016/j.jip.2010.06.012. Epub 2010 Jul 1.

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.

MeSH terms

  • Animals
  • Bees / virology*
  • DNA-Directed RNA Polymerases / genetics
  • Dicistroviridae / genetics*
  • Insect Viruses / classification
  • Insect Viruses / genetics
  • Insect Viruses / isolation & purification
  • Picornaviridae / genetics
  • Picornaviridae Infections / diagnosis
  • Picornaviridae Infections / genetics
  • Picornaviridae Infections / veterinary*
  • RNA, Viral / analysis
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary*
  • Virus Diseases / diagnosis
  • Virus Diseases / genetics
  • Virus Diseases / veterinary*

Substances

  • RNA, Viral
  • DNA-Directed RNA Polymerases