期刊
VIRUS RESEARCH
卷 211, 期 -, 页码 17-24出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.virusres.2015.09.015
关键词
West Nile virus; Reporter virus; Antiviral drug discovery; Neutralizing antibody; Gaussia luciferase
类别
资金
- National Basic Research Program of China [2012CB518904]
- National Natural Science Foundation of China [31170158]
West Nile virus (WNV) is a neurotropic human pathogen that has caused increasing infected cases over recent years. There is currently no licensed vaccine or effective drug for prevention and treatment of WNV infection in humans. To facilitate antiviral drug discovery and neutralizing antibody detection, a WNV cDNA clone containing a luciferase reporter gene was constructed through incorporating Gaussia luciferase (Gluc) gene within the capsid-coding region of WNV genome. Transfection of BHK-21 cells with the cDNA clone-derived RNA generated luciferase reporter WNV (WNV-Gluc) and the stable WNV-Gluc with high titers (>10(7) PFU/ml) was obtained through plaque purification. Luciferase activity was used to effectively quantify the viral production of WNV-Gluc. Using the reporter virus WNV-Gluc, we developed a luciferase based assay in a 12-well format for evaluating neutralizing antibodies. The reporter virus could be a powerful tool for epidemiological investigation of WNV, vaccine evaluation, antiviral drug screening, and the study of WNV replication and pathogenesis. (C) 2015 Elsevier B.V. All rights reserved.
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