Cargando…

Recovery of NanoLuc Luciferase-Tagged Canine Distemper Virus for Facilitating Rapid Screening of Antivirals in vitro

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 rec...

Descripción completa

Detalles Bibliográficos
Autores principales: Liu, Fuxiao, Wang, Qianqian, Huang, Yilan, Wang, Ning, Zhang, Youming, Shan, Hu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7758346/
https://www.ncbi.nlm.nih.gov/pubmed/33363240
http://dx.doi.org/10.3389/fvets.2020.600796
_version_ 1783626922593353728
author Liu, Fuxiao
Wang, Qianqian
Huang, Yilan
Wang, Ning
Zhang, Youming
Shan, Hu
author_facet Liu, Fuxiao
Wang, Qianqian
Huang, Yilan
Wang, Ning
Zhang, Youming
Shan, Hu
author_sort Liu, Fuxiao
collection PubMed
description 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.
format Online
Article
Text
id pubmed-7758346
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-77583462020-12-25 Recovery of NanoLuc Luciferase-Tagged Canine Distemper Virus for Facilitating Rapid Screening of Antivirals in vitro Liu, Fuxiao Wang, Qianqian Huang, Yilan Wang, Ning Zhang, Youming Shan, Hu Front Vet Sci Veterinary Science 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. Frontiers Media S.A. 2020-12-10 /pmc/articles/PMC7758346/ /pubmed/33363240 http://dx.doi.org/10.3389/fvets.2020.600796 Text en Copyright © 2020 Liu, Wang, Huang, Wang, Zhang and Shan. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Liu, Fuxiao
Wang, Qianqian
Huang, Yilan
Wang, Ning
Zhang, Youming
Shan, Hu
Recovery of NanoLuc Luciferase-Tagged Canine Distemper Virus for Facilitating Rapid Screening of Antivirals in vitro
title Recovery of NanoLuc Luciferase-Tagged Canine Distemper Virus for Facilitating Rapid Screening of Antivirals in vitro
title_full Recovery of NanoLuc Luciferase-Tagged Canine Distemper Virus for Facilitating Rapid Screening of Antivirals in vitro
title_fullStr Recovery of NanoLuc Luciferase-Tagged Canine Distemper Virus for Facilitating Rapid Screening of Antivirals in vitro
title_full_unstemmed Recovery of NanoLuc Luciferase-Tagged Canine Distemper Virus for Facilitating Rapid Screening of Antivirals in vitro
title_short Recovery of NanoLuc Luciferase-Tagged Canine Distemper Virus for Facilitating Rapid Screening of Antivirals in vitro
title_sort recovery of nanoluc luciferase-tagged canine distemper virus for facilitating rapid screening of antivirals in vitro
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7758346/
https://www.ncbi.nlm.nih.gov/pubmed/33363240
http://dx.doi.org/10.3389/fvets.2020.600796
work_keys_str_mv AT liufuxiao recoveryofnanolucluciferasetaggedcaninedistempervirusforfacilitatingrapidscreeningofantiviralsinvitro
AT wangqianqian recoveryofnanolucluciferasetaggedcaninedistempervirusforfacilitatingrapidscreeningofantiviralsinvitro
AT huangyilan recoveryofnanolucluciferasetaggedcaninedistempervirusforfacilitatingrapidscreeningofantiviralsinvitro
AT wangning recoveryofnanolucluciferasetaggedcaninedistempervirusforfacilitatingrapidscreeningofantiviralsinvitro
AT zhangyouming recoveryofnanolucluciferasetaggedcaninedistempervirusforfacilitatingrapidscreeningofantiviralsinvitro
AT shanhu recoveryofnanolucluciferasetaggedcaninedistempervirusforfacilitatingrapidscreeningofantiviralsinvitro