基于重组酶介导扩增-侧流层析试纸条的褐飞虱快速鉴定方法

【目的】针对测报灯下褐飞虱鉴定费时费力这一问题,建立褐飞虱快速定性鉴定技术。【方法】以褐飞虱、伪褐飞虱及拟褐飞虱为材料,筛选褐飞虱特异性引物对,优化基于重组酶介导扩增和侧流层析试纸条检测方法的快速鉴定体系(RAA-LFD),并分析其温度及模板鲁棒性。【结果】本研究建立的RAA-LFD方法操作简单、速度快、不依赖于任何精密仪器、体温可触发扩增反应,且对粗组织液模板具有很好的鲁棒性。盲样检测结果表明,56个样本从样本获取到结果输出的整个过程可在45 min内完成,准确率可达100%。【结论】本研究建立的褐飞虱RAA-LFD鉴定技术可以用于褐飞虱及其近似种的快速区分,适用于基层植保站或田间地头对样本的现场即时检测。

A Rapid Detection Assay of Nilaparvata lugens Based on Recombinase Aided Amplification-lateral Flow Dipstick Technology

【Objective】To solve the time- and labor-costing problem for accurate identification of Nilarpavata lugens trapped by light, we aim to establish a fast and convenient qualitative identification method.【Method】At first, we selected a couple of primers which are species specific for N. lugens. Then, we established an identification method based on Rapid Aided Amplification-Lateral Flow Dipstick (RAA-LFD) technology. Finally, we analyzed the temperature and template robustness of this method. 【Result】The RAA-LFD method developed in the present study is convenient and fast, and it does not depend on any sophisticated instrument. Furthermore, this method can amplify effectively with crude tissue fluid as template, and it can be triggered by body temperature. The entire process for 56 specimens, including the crude tissue liquid preparation, isothermal amplification, and LFD detection, can be completed in 45 min with an accuracy of 100%. 【Conclusion】This RAA-LFD method is a time-saving and easy-to-apply molecular diagnostic method for N.lugens identification from its two sibling species. It is really suitable for Point of Care Test (POCT) in plant protection station or in field.