Diagnosis of the black queen cell virus (BQCV) by loop-mediated isothermal amplification
Author of the article:ZHUANG Ming-Liang** LI Jiang-Hong CHEN Da-Fu LIANG Qin***
Author's Workplace：College of Bee Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China
Key Words：black queen cell virus, loop-mediated isothermal amplification, detection, diagnosis
Abstract：[Objectives] To establish a simple, fast and accurate method to detect black queen cell virus (BQCV) using loop-mediated isothermal amplification (LAMP), which provides an experimental data for controlling the disease. [Methods] Four primers based on six conserved regions of the BQCV gene sequence were used to develop optimal LAMP amplification conditions and the resultant LAMP amplification results compared those of conventional PCR (polymerase chain reaction). [Results] The LAMP method could detect viral concentrations as low as 86 fg DNA, which is 100 times more sensitive than PCR. Clinical results show that LAMP can be used to detect BQCV in either Italian (Apis mellifera ligustica) or Chinese bees (Apis cerana cerana), with a true positive detection ratio 10%-20% higher than that of PCR. [Conclusion] The newly developed LAMP test is a practical and valuable method for detecting and controlling BQCV in honeybees.