Sacbrood virus (SBV) is one of the most serious honeybee viruses. The virus causes failure to pupate and death in both larvae and adult bees. Recently, Korean Sacbrood virus (KSBV) caused a great loss in Korean honeybee (Apis cerana) colonies. Although KSBV shows high homology with SBV strains, it has unique motifs and causes different symptoms. Therefore, a simple, sensitive and specific method for detecting KSBV is needed urgently. In this study a reverse transcription loop-mediated isothermal amplification (RT-LAMP) and a novel micro PCR-based detection method, termed ultra-rapid real-time PCR (URRT-PCR) were applied for rapid detection for korean sacbrood virus (KSBV) from honeybees (Apis cerana) infected with SBV in Korea. The LAMP could be detect the virus in RT-LAMP reactions containing 102copies of pBX-KSBV within 30 min, which was 10 times more sensitive than a RT-PCR assay. The URRT-PCR showed high sensitivities which were able to detect 10 copies in the standard assays. In the application of URRT-PCR detection to an KSBV-infected honeybee, the shortest detection time was 10 min 12 sec, including reverse transcription. In addition, these methods could be distinguished between KSBV and other closely-related SBV strains, These rapid methods were rapid molecular-based diagnostic tools and useful tool for the rapid and sensitive diagnosis of KSBV infection of honeybees.

• Mi-Sun Yoo(Center for Honeybee Disease Control, Parasite and Insect Disease Laboratory, Bacterial Disease Division, Animal and Plant Quarantine Agency.)
• Young-Ha Kim(Center for Honeybee Disease Control, Parasite and Insect Disease Laboratory, Bacterial Disease Division, Animal and Plant Quarantine Agency.)
• Nam-Hee Kim(Center for Honeybee Disease Control, Parasite and Insect Disease Laboratory, Bacterial Disease Division, Animal and Plant Quarantine Agency.)
• Ha-Na Jung(Center for Honeybee Disease Control, Parasite and Insect Disease Laboratory, Bacterial Disease Division, Animal and Plant Quarantine Agency.)
• Kondreddy Eswar Reddy(Center for Honeybee Disease Control, Parasite and Insect Disease Laboratory, Bacterial Disease Division, Animal and Plant Quarantine Agency.)
• Suk-Chan Jung(Center for Honeybee Disease Control, Parasite and Insect Disease Laboratory, Bacterial Disease Division, Animal and Plant Quarantine Agency.)
• Seung-Won Kang(Center for Honeybee Disease Control, Parasite and Insect Disease Laboratory, Bacterial Disease Division, Animal and Plant Quarantine Agency.)