应用环介导等温扩增技术（LAMP）检测蜜蜂黑蜂王台病毒的研究
Diagnosis of the black queen cell virus (BQCV) by loop-mediated isothermal amplification
庄明亮** 李江红 陈大福 梁 勤***
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DOI：10.7679/j.issn.2095-1353.2014.192
作者单位：福建农林大学蜂学学院，福州 350002
中文关键词：黑蜂王台病毒，环介导等温扩增，检测，诊断
英文关键词：black queen cell virus, loop-mediated isothermal amplification, detection, diagnosis
中文摘要：
【目的】 本文旨在建立用于临床检测黑蜂王台病毒（Black queen cell virus，BQCV）的环介导等温扩增技术（Loop-mediated isothermal amplification，LAMP），为该疾病的检测和防控提供技术支撑。【方法】 根据BQCV基因保守序列设计4条特异性引物，探究LAMP扩增的最优条件，并与常规的PCR（Polymerase chain reaction）检测方法进行比较。【结果】 建立的LAMP方法特异性好，检测下限为86 fg， 灵敏度比PCR高100倍。临床检测显示其对意大利蜜蜂Apis mellifera ligustica和中华蜜蜂Apis cerana cerana均准确有效，且检出率比普通PCR法高10%~20%。【结论】 建立的针对BQCV的LAMP检测方法为养蜂生产第一线检测和预防BQCV提供了技术支持，有一定的应用价值。
英文摘要：[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.