首页|A high throughput antiviral screening platform for alphaviruses based on Semliki Forest virus expressing eGFP reporter gene
A high throughput antiviral screening platform for alphaviruses based on Semliki Forest virus expressing eGFP reporter gene
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Alphaviruses,which contain a variety of mosquito-borne pathogens,are important pathogens of emerging/re-emerging infectious diseases and potential biological weapons.Currently,no specific antiviral drugs are avail-able for the treatment of alphaviruses infection.For most highly pathogenic alphaviruses are classified as risk group-3 agents,the requirement of biosafety level 3(BSL-3)facilities limits the live virus-based antiviral study.To facilitate the antiviral development of alphaviruses,we developed a high throughput screening(HTS)platform based on a recombinant Semliki Forest virus(SFV)which can be manipulated in BSL-2 laboratory.Using the reverse genetics approach,the recombinant SFV and SFV reporter virus expressing eGFP(SFV-eGFP)were suc-cessfully rescued.The SFV-eGFP reporter virus exhibited robust eGFP expression and remained relatively stable after four passages in BHK-21 cells.Using a broad-spectrum alphavirus inhibitor ribavirin,we demonstrated that the SFV-eGFP can be used as an effective tool for antiviral study.The SFV-eGFP reporter virus-based HTS assay in a 96-well format was then established and optimized with a robust Z'score.A section of reference compounds that inhibit highly pathogenic alphaviruses were used to validate that the SFV-eGFP reporter virus-based HTS assay enables rapid screening of potent broad-spectrum inhibitors of alphaviruses.This assay provides a safe and convenient platform for antiviral study of alphaviruses.
Yu-Jia Shi、Jia-Qi Li、Hong-Qing Zhang、Cheng-Lin Deng、Qin-Xuan Zhu、Bo Zhang、Xiao-Dan Li
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Hunan Normal University,School of Medicine,Changsha,410081,China
Key Laboratory of Special Pathogens and Biosafety,Wuhan Institute of Virology,Center for Biosafety Mega-Science,Chinese Academy of Sciences,Wuhan,430071,China
University of Chinese Academy of Sciences,Beijing,100049,China
Creative Research Group Program of Natural Science Foundation of Hubei ProvinceNational Natural Science Foundation of China