Polymerase Chain Reaction detection of deformed wing virus (DWV) in Apis mellifera and Varroa destructor
Tentcheva, Diana | Gauthier, Laurent | Jouve, Sandrine | Canabady-Rochelle, Laetitia | Dainat, Benjamin | Cousserans, Francois | Colin, Marc Edouard | Ball, Brenda V. | Bergoin, Max
We have developed a specific assay for the detection of deformed wing virus (DWV) in Apis mellifera L. and Varroa destructor based on the reverse transcriptase polymerase chain reaction (RT-PCR) technology. Primers were designed from the sequence of a 4700 nucleotides cDNA fragment located in the 3'-end of the DWV genome. This fragment encodes a single open reading frame of 1565 amino acids showing similarity to viral RNA dependent RNA polymerase consensus motif. RT-PCR assays from DWV infected individual mite or bee produced a 395 nucleotide DNA fragment clearly identifiable by agarose gel electrophoresis. The signal in bees having deformed wings was significantly higher than in normal ones. A search for DWV in 40 colonies showed that DWV is broadly distributed in bee colonies and mites. As an average, greater virus prevalence of virus was detected in bees collected in autumn compared to bees collected in spring or during the summer period
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