title: Development of Neutralization Assay Using an eGFP Chikungunya Virus.
abstract: Chikungunya virus (CHIKV), a member of the Alphavirus genus, is an important human emerging/re-emerging pathogen. Currently, there are no effective antiviral drugs or vaccines against CHIKV infection. Herein, we construct an infectious clone of CHIKV and an eGFP reporter CHIKV (eGFP-CHIKV) with an isolated strain (assigned to Asian lineage) from CHIKV-infected patients. The eGFP-CHIKV reporter virus allows for direct visualization of viral replication through the levels of eGFP expression. Using a known CHIKV inhibitor, ribavirin, we confirmed that the eGFP-CHIKV reporter virus could be used to identify inhibitors against CHIKV. Importantly, we developed a novel and reliable eGFP-CHIKV reporter virus-based neutralization assay that could be used for rapid screening neutralizing antibodies against CHIKV.
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. Dengchenglin29@126.com.
authors.forename: Cheng-Lin
authors.initials: CL
authors.lastname: Deng
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. luckyliu_110@163.com.
authors.forename: Si-Qing
authors.initials: SQ
authors.lastname: Liu
authors.affiliation: Ningbo International Travel Healthcare Center, Ningbo 315012, China. zhoudg@nbciq.gov.cn.
authors.forename: Dong-Gen
authors.initials: DG
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. Xulinlin.2007yahoo@163.com.
authors.forename: Lin-Lin
authors.initials: LL
authors.lastname: Xu
authors.affiliation: School of Medicine, Hunan Normal University, Changsha 410000, China. lxd@live.cn.
authors.forename: Xiao-Dan
authors.initials: XD
authors.lastname: Li
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. zhangpantao123@163.com.
authors.forename: Pan-Tao
authors.initials: PT
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. lipenghui101@126.com.
authors.forename: Peng-Hui
authors.initials: PH
authors.lastname: Li
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. yehq@wh.iov.cn.
authors.forename: Han-Qing
authors.initials: HQ
authors.lastname: Ye
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. hpwei@wh.iov.cn.
authors.forename: Hong-Ping
authors.initials: HP
authors.lastname: Wei
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. yzm@wh.iov.cn.
authors.forename: Zhi-Ming
authors.initials: ZM
authors.lastname: Yuan
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China. chengfeng_qin@126.com.
authors.forename: Cheng-Feng
authors.initials: CF
authors.lastname: Qin
authors.affiliation: Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China. zhangbo@wh.iov.cn.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D002460:Cell Line; D002646:Chikungunya virus / Q000235:genetics / Q000276:immunology* / Q000502:physiology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D008403:Mass Screening / Q000379:methods; D009500:Neutralization Tests / Q000379:methods*; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods*; D014779:Virus Replication
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Chikungunya virus; antiviral; neutralization antibody; reporter virus