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.

AlphavirusSemliki Forest virusReporter virusHigh throughput screeningInhibitor

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

2022CFA02181702005

2023

中国病毒学
中国科学院武汉病毒研究所,中国微生物学会

中国病毒学

CSTPCDCSCD
影响因子:0.393
ISSN:1674-0769
年,卷(期):2023.38(4)
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