Abstract
Canine distemper virus (CDV), belonging to the genus Morbillivirus in the family Paramyxoviridae, 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 in vitro. 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 in vitro. 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) via analysis of luciferase activity, instead of via 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.
Highlights
Canine distemper (CD) is a highly contagious disease, affecting a wide variety of domestic and wild carnivores [1]
Canine distemper virus (CDV) is an ideal vector for expressing various foreign proteins, including green fluorescence protein (GFP) [21], red fluorescence protein [22], firefly luciferase [10], interleukin-7 [23], interleukin-18 [24], and rabies virus glycoprotein [25]
The rCDV-NanoLuc® luciferase (NLuc) could be readily rescued from its cDNA clone, because of bright luminescence that was visible at 72 hpt or even earlier
Summary
Canine distemper (CD) is a highly contagious disease, affecting a wide variety of domestic and wild carnivores [1]. The etiological agent of CD is canine distemper virus (CDV), which has been renamed canine morbillivirus, according to the latest virus taxonomy of the International Committee on Taxonomy of Viruses. Available CDV vaccines can confer protective immunity in vivo, whereas outbreaks of CD are still frequently reported in domestic and wild animals [3]. Both practitioners and dog owners have a strong demand for effective anti-CDV drugs, still unavailable to date. Dogs with early-stage CDV infection can be passively immunized with
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