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埃博拉病毒属于丝状病毒家族,可引起人类恶性传染病。本文旨在建立可用于高通量筛选评价丝状病毒进入抑制剂的细胞水平重组病毒模型。基于重组病毒技术,共建立了三株丝状重组病毒模型,分别是扎伊尔型埃博拉重组病毒、苏丹型埃博拉重组病毒和马尔堡重组病毒。该模型以丝状病毒表面唯一负责病毒进入的糖蛋白为病毒表面蛋白,包裹带有荧光素酶报告基因的HIV内核,组装为丝状重组病毒颗粒。当病毒感染细胞后,病毒感染程度可通过检测细胞中荧光素酶活性获得。此模型靶点明确,安全性好,当进行丝状病毒进入抑制剂筛选时,可通过与水泡性口膜炎重组病毒联用判断化合物的特异性。应用此模型对文献报道的丝状病毒进入抑制剂进行评价,数据与文献报道相符,模型构建成功。丝状重组病毒进入模型的建立对于抗丝状病毒药物的快速筛选和研发有重要意义。
Ebola virus belongs to the family of filariasis, can cause human malignant infectious diseases. This article aims to establish a cellular level recombinant virus model that can be used to screen for high-throughput screening of inhibitors of filamentous virus entry. Based on the recombinant virus technology, a total of three filamentous recombinant virus models have been established, which are ZE-Ebola virus, Sudan-type Ebola virus and Marburg virus. In this model, the only glycoprotein responsible for viral entry on the surface of the filamentous virus is the viral surface protein, which is wrapped in the HIV core with the luciferase reporter gene and assembled into filamentous recombinant virions. After the virus infects the cells, the degree of viral infection can be obtained by measuring the luciferase activity in the cells. The target of this model is clear and safe, and when screening for inhibitors of filamentous virus entry, the specificity of the compound can be judged by the combination with vesicular stomatitis virus. The model was used to evaluate the reported inhibitors of filamentous virus entering into the literature. The data were consistent with those reported in the literature and the model was successfully constructed. The establishment of a filamentous recombinant virus entry model is of great importance for the rapid screening and development of anti-filamentous drugs.