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http://dx.doi.org/10.7845/kjm.2013.3052

Detection of a Microsporidium, Nosema ceranae, from Field Population of the Bumblebee, Bombus terrestris, via Quantitative Real-Time PCR  

Lee, Dae-Weon (Department of Biology, Kyungsung University)
Publication Information
Korean Journal of Microbiology / v.49, no.3, 2013 , pp. 270-274 More about this Journal
Abstract
The bumblebee, Bombus terrestris, has played an important role as one of the alternative pollinators since the outbreak of honeybee collapse disorder. Recently, pathogens and parasites such as viruses, bacteria and mites, which affect the life span and fecundity of their host, have been discovered in B. terristris. In order to detect the microsporidian pathogen, Nosema spp. in the field populations of B. terristris, we collected adults and isolated their genomic DNA for diagnostic PCR. The PCR primers specific for Nosema spp. were newly designed and applied to gene amplification for cloning. Only small subunit ribosomal RNA (SSU rRNA) gene of N. ceranae was successfully amplified among examined genes and sequenced, which indicates that N. ceranae mainly infects the examined field population of B. terristris. To detect of SSU rRNA gene, two regions of SSU rRNA gene were selected by primary PCR analysis and further analyzed in quantitative real-time PCR (qRT-PCR). The qRT-PCR analysis demonstrated that SSU rRNA of N. ceranae was detected at concentration as low as $0.85ng/{\mu}l$ genomic DNA. This result suggests that the detection via qRT-PCR can be applied for the rapid and sensitive diagnosis of N. ceranae infection in the field population as well as risk assessment of B. terristris.
Keywords
Bombus terrestris; Nosema ceranae; diagnosis; quantitative real-time PCR; risk assessment;
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