通过逆转录环介导等温扩增技术快速灵敏地检测百合无症状病毒
Rapid and sensitive detection of Lily symptomless virus by reverse transcription loop-mediated isothermal amplification.
作者信息
He Xiangfeng, Xue Fei, Xu Shufa, Wang Wenhe
机构信息
Beijing Engineering Research Center of Rural Landscape Planning and Design, College of Landscape Architecture, Beijing University of Agriculture, Beijing 102206, China; Beijing Collaborative Innovation Center for Eco-Environmental Improvement with Forestry and Fruit Trees, Beijing 102206, China.
Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China.
出版信息
J Virol Methods. 2016 Dec;238:38-41. doi: 10.1016/j.jviromet.2016.10.003. Epub 2016 Oct 11.
Lily symptomless virus (LSV) is one of the most prevalent viruses that infect lily plants worldwide. A rapid and sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of LSV, using two primer pairs that specifically amplified the conserved sequence of LSV coat protein. The optimum reaction conditions were as follows: 4mM MgCl and 0.8M betaine with incubation at 64°C for 30min. The limit of detection of LSV from infected lily leaves was 10-fold higher for RT-LAMP than for conventional RT-PCR. Moreover, RT-LAMP detected LSV in not only symptomatic, but also in symptomless tissues of infected plants. These findings indicate that our RT-LAMP method for LSV can serve as a low-cost, simple, and rapid alternative to conventional detection assays.
百合无症状病毒（LSV）是全球感染百合植物最普遍的病毒之一。开发了一种快速灵敏的逆转录环介导等温扩增（RT-LAMP）检测方法用于检测LSV，使用两对特异性扩增LSV外壳蛋白保守序列的引物。最佳反应条件如下：4mM MgCl和0.8M甜菜碱，在64°C孵育30分钟。从受感染的百合叶片中检测LSV时，RT-LAMP的检测限比传统RT-PCR高10倍。此外，RT-LAMP不仅能在受感染植物的有症状组织中检测到LSV，还能在无症状组织中检测到。这些结果表明，我们用于检测LSV的RT-LAMP方法可作为一种低成本、简单且快速的传统检测方法的替代方法。
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