title: Generation of a recombinant West Nile virus stably expressing the Gaussia luciferase for neutralization assay.
abstract: 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.
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Science, Wuhan 430071, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Pan-Tao
authors.initials: PT
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Science, Wuhan 430071, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Shan
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Science, Wuhan 430071, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Xiao-Dan
authors.initials: XD
authors.lastname: Li
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Science, Wuhan 430071, China.
authors.forename: Si-Qing
authors.initials: SQ
authors.lastname: Liu
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Science, Wuhan 430071, China.
authors.forename: Cheng-Lin
authors.initials: CL
authors.lastname: Deng
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Science, Wuhan 430071, China.
authors.forename: Han-Qing
authors.initials: HQ
authors.lastname: Ye
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Science, Wuhan 430071, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Bao-Di
authors.initials: BD
authors.lastname: Shang
authors.affiliation: Novartis Institute for Tropical Diseases, Singapore.
authors.forename: Pei-Yong
authors.initials: PY
authors.lastname: Shi
authors.affiliation: Beijing Institute of Basic Medical Sciences, Beijing 100850, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Lv
authors.affiliation: Beijing Institute of Basic Medical Sciences, Beijing 100850, China.
authors.forename: Bei-Fen
authors.initials: BF
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Cheng-Feng
authors.initials: CF
authors.lastname: Qin
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Science, Wuhan 430071, China. Electronic address: zhangbo@wh.iov.cn.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000032:analysis* / Q000276:immunology; D000914:Antibodies, Viral / Q000032:analysis / Q000276:immunology; D033342:Copepoda; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D009500:Neutralization Tests / Q000295:instrumentation / Q000379:methods*; D014901:West Nile Fever / Q000276:immunology / Q000821:virology; D014902:West Nile virus / Q000235:genetics / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D008156:Luciferases
keywords: Antiviral drug discovery; Gaussia luciferase; Neutralizing antibody; Reporter virus; West Nile virus