Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, China; Guangdong Silda Pupa and Silk Inc., Shaoguan, China.
Collage of Life Science, Fudan University, Shanghai 200032, China; Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China.
J Invertebr Pathol. 2019 May;163:75-81. doi: 10.1016/j.jip.2019.03.003. Epub 2019 Mar 9.
Bombyx mori bidensovirus (BmBDV), is the only bipartite single-stranded DNA (ssDNA) insect virus reported to date. BmBDV causes fatal flacherie disease in silkworm, resulting in large economic losses to sericulture. We developed a loop-mediated isothermal amplification with lateral flow dipstick (LAMP-LFD) method that can successfully and rapidly detect BmBDV DNA with a low limit of detection (5 fg, 400 copies of the BmBDV genome). The method was evaluated and improved for direct field diagnosis using silkworm faeces. In the field, the actual limit of detection was ∼50 fg (4000 copies of the BmBDV genome). The results demonstrated that, compared with traditional methods for BmBDV detection, our new LAMP-LFD method was much more rapid, sensitive and cost-effective, with less dependence on equipment, making it easier to use. The method has potential to be translated into a new diagnostic product for field applications in the sericulture industry.
家蚕双瓣轮状病毒（BmBDV）是迄今为止报道的唯一一种具有二分体的单链 DNA（ssDNA）昆虫病毒。BmBDV 可引起家蚕致命的软化病，给蚕桑业造成巨大的经济损失。我们开发了一种环介导等温扩增与侧向流试纸条（LAMP-LFD）方法，该方法可以成功且快速地检测 BmBDV DNA，检测限低至 5 fg（400 个 BmBDV 基因组拷贝）。该方法经过评估和改进，可直接用于田间诊断家蚕粪便。在田间条件下，实际检测限约为 50 fg（4000 个 BmBDV 基因组拷贝）。结果表明，与 BmBDV 检测的传统方法相比，我们的新型 LAMP-LFD 方法更加快速、灵敏和具有成本效益，对设备的依赖性更小，使用起来更加方便。该方法有望转化为一种新的诊断产品，用于蚕桑业的现场应用。
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