A high throughput antiviral screening platform for alphaviruses based on Semliki Forest virus expressing eGFP reporter gene  

作　　者：Yu-Jia Shi Jia-Qi Li Hong-Qing Zhang Cheng-Lin Deng Qin-Xuan Zhu Bo Zhang Xiao-Dan Li 

机构地区：[1]Hunan Normal University,School of Medicine,Changsha,410081,China [2]Key Laboratory of Special Pathogens and Biosafety,Wuhan Institute of Virology,Center for Biosafety Mega-Science,Chinese Academy of Sciences,Wuhan,430071,China [3]University of Chinese Academy of Sciences,Beijing,100049,China

出　　处：《Virologica Sinica》2023年第4期585-594,共10页中国病毒学（英文版）

基　　金：supported by the Creative Research Group Program of Natural Science Foundation of Hubei Province (2022CFA021);National Natural Science Foundation of China (81702005).

摘　　要：Alphaviruses,which contain a variety of mosquito-borne pathogens,are important pathogens of emerging/reemerging infectious diseases and potential biological weapons.Currently,no specific antiviral drugs are available 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 successfully 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 Z0 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.

关 键 词：ALPHAVIRUS Semliki Forest virus Reporter virus High throughput screening INHIBITOR 

分 类 号：R965.1[医药卫生—药理学]