Prompt Diagnosis of Bombyx mori Nucleopoyhedrosis Virus by Loop-mediated Isothermal Amplification

Author of the article:TANG Fen-Fen TANG FenFenYANG WeiKeSHAO YuLanXIE DaoYanLIAO PengFeiYANG HaiBAI XingRong

Author's Workplace：Institute of Sericulture and Apiculture，Yunnan Academy of Agricultural Sciences

Key Words：Bombyx mori nucleopoyhedrosis virus (BmNPV), Loop-mediated isothermal amplification(LAMP), Diognosis, Sericulture

Abstract：Bombyx mori nucleopoyhedrosis virus (BmNPV) infects silkworm severely and leads to economic damage in sericulture. In this study, a robust and simple Loop-mediated isothermal amplification （LAMP） assay for rapid detection of BmNPV was developed，which is the first suitable for potential diagnosis and helping to monitor BmNPV infections in the sericulture. A set of six specific primers was designed to recognize six distinct genomic sequences of pe38 from BmNPV. The entire assay duration was recorded at 25min under isothermal condition at 63℃.The amplified products were analyzed by electrophoresis and visual judgment by the SYBR Green I dyeing. The results showed that LAMP assay was 100-fold more sensitive than the routine PCR assay, with a detection limit of 21 copies per reaction. The amount of viral DNA in hemolymph sampled from the larvae infected at different time points was also invested by PCR and LAMP