用于促进抗病毒药物快速筛选的纳米荧光素酶标记犬瘟热病毒的复苏
Recovery of NanoLuc Luciferase-Tagged Canine Distemper Virus for Facilitating Rapid Screening of Antivirals .
作者信息
Liu Fuxiao, Wang Qianqian, Huang Yilan, Wang Ning, Zhang Youming, Shan Hu
机构信息
College of Veterinary Medicine, Qingdao Agricultural University, Qingdao, China.
State Key Laboratory of Microbial Technology, Shandong University, Qingdao, China.
出版信息
Front Vet Sci. 2020 Dec 10;7:600796. doi: 10.3389/fvets.2020.600796. eCollection 2020.
Canine distemper virus (CDV), belonging to the genus in the family , is a highly contagious pathogen, affecting various domestic, and wild carnivores. Conventional methods are too cumbersome to be used for high-throughput screening of anti-CDV drugs. In this study, a recombinant CDV was rescued using reverse genetics for facilitating screening of anti-CDV drug . The recombinant CDV could stably express the NanoLuc® luciferase (NLuc), a novel enzyme that was smaller and "brighter" than others. The intensity of NLuc-catalyzed luminescence reaction indirectly reflected the anti-CDV effect of a certain drug, due to a positive correlation between NLuc expression and virus propagation . Based on such a characteristic feature, the recombinant CDV was used for anti-CDV assays on four drugs (ribavirin, moroxydine hydrochloride, 1-adamantylamine hydrochloride, and tea polyphenol) analysis of luciferase activity, instead of conventional methods. The result showed that out of these four drugs, only the ribavirin exhibited a detectable anti-CDV effect. The NLuc-tagged CDV would be a rapid tool for high-throughput screening of anti-CDV drugs.
犬瘟热病毒（CDV）属于 科 属，是一种高度传染性的病原体，可感染各种家养和野生食肉动物。传统方法过于繁琐，无法用于高通量筛选抗CDV药物。在本研究中，利用反向遗传学拯救了一种重组CDV，以促进抗CDV药物的筛选。重组CDV能够稳定表达纳米荧光素酶（NLuc），这是一种比其他酶更小且“更亮”的新型酶。由于NLuc表达与病毒增殖之间呈正相关，NLuc催化的发光反应强度间接反映了某种药物的抗CDV效果。基于这一特征，使用重组CDV对四种药物（利巴韦林、盐酸吗啉胍、盐酸金刚烷胺和茶多酚）进行抗CDV测定，并分析荧光素酶活性，而不是使用传统方法。结果表明，在这四种药物中，只有利巴韦林表现出可检测到的抗CDV效果。带有NLuc标签的CDV将成为高通量筛选抗CDV药物的快速工具。
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