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KMID : 0364820130490030270
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Korean Journal of Microbiology
2013 Volume.49 No. 3 p.270 ~ p.274
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Detection of a Microsporidium, Nosema ceranae, from Field Population of the Bumblebee, Bombus terrestris, via Quantitative Real-Time PCR
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Lee Dae-Weon
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Abstract
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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 0.85 ng/μl 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.
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KEYWORD
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Bombus terrestris, Nosema ceranae, diagnosis, quantitative real-time PCR, risk assessment
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