Abstract:

 To establish a rapid and effective quarantine method of the honeybee sacbrood virus disease, we developed a real-time RT-PCR assay for detection of bee sacbrood disease using TaqMan probes. A pair of specific primers and a probe used in this assay were designed based on a highly conserved region in sacbrood bee virus. The assay was shown to be sensitive, detecting less than 1.0×10² copies/μL, and specific for the detection of sacbrood bee virus. Cross-reaction with acute bee paralysis virus, deformed wing virus and black queen cell virus was not observed. The coefficient of variation in the stability experiments was 1.6%. A reliable diagnostic result can be obtained just within 4 h. The assay proved to be a rapid, sensitive, specific and repetitive method for rapid detection of sacbrood bee virus from honeybee and honeybee products in quarantine.

Key words: Honeybee, sacbrood virus (SBV), TaqMan probe, real-time PCR, quarantine