Isorce Nathalie, Fasel Nicolas
Department of Biochemistry, University of Lausanne, Epalinges, Switzerland.
Bio Protoc. 2020 May 5;10(9):e3598. doi: 10.21769/BioProtoc.3598.
Many RNA viruses are found in protozoan parasites. They can be responsible for more serious pathology or treatment failure. For the detection of viral double-stranded RNA (dsRNA), sequence-dependent and -independent methods are available, such as quantitative real-time PCR and immunofluorescence, dot blot, ELISA or sequencing. The technique presented here is sequence-independent and is well detailed in the following protocol, taking the example of RNA virus (LRV) in ) species. To summarise, the protocol is divided into four major steps: RNA extraction from the parasites, RNA purification, enzymatic digestions with DNase I and Nuclease S1, and visualization by gel electrophoresis. This method can be used to detect other viral dsRNA in other parasites. It provides an additional tool, complementary to other techniques previously cited and it is easy and quite fast to achieve.
许多RNA病毒存在于原生动物寄生虫中。它们可能导致更严重的病理状况或治疗失败。对于病毒双链RNA（dsRNA）的检测，有依赖序列和不依赖序列的方法，如定量实时PCR、免疫荧光、斑点印迹、ELISA或测序。本文介绍的技术是不依赖序列的，以下方案将以 种中的RNA病毒（LRV）为例进行详细说明。总之，该方案分为四个主要步骤：从寄生虫中提取RNA、RNA纯化、用DNase I和核酸酶S1进行酶切消化以及通过凝胶电泳进行可视化。该方法可用于检测其他寄生虫中的其他病毒dsRNA。它提供了一种额外的工具，是对先前引用的其他技术的补充，并且操作简便、速度较快。
