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Applied and Environmental Microbiology, January 2005, p. 436-441, Vol. 71, No. 1
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.1.436-441.2005

Quantitative Real-Time Reverse Transcription-PCR Analysis of Deformed Wing Virus Infection in the Honeybee (Apis mellifera L.)

Y. P. Chen,1* J. A. Higgins,2 and M. F. Feldlaufer1

Bee Research Laboratory,1 Environmental Microbial Safety Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Beltsville, Maryland2

Received 14 May 2004/ Accepted 4 August 2004

Deformed wing virus (DWV) can cause wing deformity and premature death in adult honeybees, although like many other bee viruses, DWV generally persists as a latent infection with no apparent symptoms. Using reverse transcription (RT)-PCR and Southern hybridization, we detected DWV in all life stages of honeybees, including adults with and without deformed wings. We also found DWV in the parasitic mite Varroa destructor, suggesting that this mite may be involved in the transmission of DWV. However, the detection of the virus in life stages not normally associated with mite parasitism (i.e., eggs and larvae) suggests that there are other modes of transmission. The levels of DWV in different life stages of bees were investigated by using TaqMan real-time quantitative RT-PCR. The amounts of virus varied significantly in these different stages, and the highest levels occurred in pupae and in adult worker bees with deformed wings. The variability in virus titer may reflect the different abilities of bees to resist DWV infection and replication. The epidemiology of DWV is discussed, and factors such as mite infestation, malnutrition, and climate are also considered.

* Corresponding author. Mailing address: Bee Research Laboratory, USDA-ARS, Bldg. 476, BARC-East, Beltsville, MD 20705. Phone: (301) 504-8749. Fax: (301) 504-8736. E-mail: chenj{at}ba.ars.usda.gov.

Applied and Environmental Microbiology, January 2005, p. 436-441, Vol. 71, No. 1
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.1.436-441.2005




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