Generation of a recombinant West Nile virus stably expressing the Gaussia luciferase for neutralization assay

Pan Tao Zhang; Chao Shan; Xiao Dan Li; Si Qing Liu; Cheng Lin Deng; Han Qing Ye; Bao Di Shang; Pei Yong Shi; Ming Lv; Bei Fen Shen; Cheng Feng Qin; Bo Zhang

doi:10.1016/j.virusres.2015.09.015

Generation of a recombinant West Nile virus stably expressing the Gaussia luciferase for neutralization assay

Pan Tao Zhang, Chao Shan, Xiao Dan Li, Si Qing Liu, Cheng Lin Deng, Han Qing Ye, Bao Di Shang, Pei Yong Shi, Ming Lv, Bei Fen Shen, Cheng Feng Qin, Bo Zhang

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

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 (>10⁷PFU/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.

Original language	English (US)
Pages (from-to)	17-24
Number of pages	8
Journal	Virus Research
Volume	211
DOIs	https://doi.org/10.1016/j.virusres.2015.09.015
State	Published - Jan 4 2016
Externally published	Yes

Keywords

Antiviral drug discovery
Gaussia luciferase
Neutralizing antibody
Reporter virus
West nile virus

ASJC Scopus subject areas

Virology
Infectious Diseases
Cancer Research

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10.1016/j.virusres.2015.09.015

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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 (>107PFU/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.",

keywords = "Antiviral drug discovery, Gaussia luciferase, Neutralizing antibody, Reporter virus, West nile virus",

author = "Zhang, {Pan Tao} and Chao Shan and Li, {Xiao Dan} and Liu, {Si Qing} and Deng, {Cheng Lin} and Ye, {Han Qing} and Shang, {Bao Di} and Shi, {Pei Yong} and Ming Lv and Shen, {Bei Fen} and Qin, {Cheng Feng} and Bo Zhang",

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year = "2016",

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doi = "10.1016/j.virusres.2015.09.015",

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T1 - Generation of a recombinant West Nile virus stably expressing the Gaussia luciferase for neutralization assay

AU - Zhang, Pan Tao

AU - Shan, Chao

AU - Li, Xiao Dan

AU - Liu, Si Qing

AU - Deng, Cheng Lin

AU - Ye, Han Qing

AU - Shang, Bao Di

AU - Shi, Pei Yong

AU - Lv, Ming

AU - Shen, Bei Fen

AU - Qin, Cheng Feng

AU - Zhang, Bo

PY - 2016/1/4

Y1 - 2016/1/4

N2 - 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 (>107PFU/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.

AB - 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 (>107PFU/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.

KW - Antiviral drug discovery

KW - Gaussia luciferase

KW - Neutralizing antibody

KW - Reporter virus

KW - West nile virus

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Generation of a recombinant West Nile virus stably expressing the Gaussia luciferase for neutralization assay

Abstract

Keywords

ASJC Scopus subject areas

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