Development of Rapid Detection Method for Honeybee Viral Diseases using the Loop-mediated Isothermal Amplification (LAMP)
2008
Yoo, M.S. (Kyonggi University, Suwon, Republic of Korea) | Cho, Y.H. (Kyonggi University, Suwon, Republic of Korea) | Kim, I.W. (Kyonggi University, Suwon, Republic of Korea) | Kang, M.H. (Kyonggi University, Suwon, Republic of Korea) | Kwon, S.H. (Chronic Inflammatory Disease Center, School of Medicine, Ajou University, Suwon, Republic of Korea) | Han, S.H. (Kyonggi University, Suwon, Republic of Korea) | Yoon, B.S. (Kyonggi University, Suwon, Republic of Korea), E-mail: bsyoon@kyonggi.ac.kr
Bee viral infections are very difficult to be identified by a macroscopic way, so an easy detection method is demanded to effectively monitor and control honeybee viral diseases in apiaries. For detection of BQCV, a BQCV-specific loop-mediated isothermal amplification (LAMP) detection method was developed. In this study, we added SybrGreen into PCR product after LAMP, visualizing amplified products from BQCV-infected honeybee samples by green fluorescent. LAMP only needs a water bath instead of sophisticate equipments, therefore this method may be very suitable for field research. In addition, two IAPV-specific primer sets (IAPV-InnerF/R and IAPV-Outer-F/R) were designed and evaluated for application of LAMP to IAPV detection. The IAPV-specific DNA fragments could be amplified less than 60 minutes by Bst DNA polymerase under an isothermal condition in the temperature range between 55℃ and 60℃. This method is expected not only to be used for BQCV and IAPV field detection but also to be applied to development of field detection method for various viral pathogens of honeybees.
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