Development and effectiveness of pseudotyped SARS-CoV-2 system as determined by neutralizing efficiency and entry inhibition test in vitro

查看全文 作　　者：Ren [1]Yang;Baoying [1]Huang;Ruhan [1]A;Wenhui [2,3]Li;Wenling [1]Wang;Yao [1]Deng;Wenjie [1]Tan

机构地区：[1]NHC Key Laboratory of Biosafety,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;[2]National Institute of Biological Sciences,No.7 Science Park Road,ZGC Life Science Park,Changping,Beijing 102206,China;[3]Tsinghua Institute of Multidisciplinary Biomedical Research,Tsinghua University,Beijing 102206,China

出　　处：《Biosafety and Health》索引2020年第2卷第4期,共6页

基　　金：support this work:The National Key Research and Development Program of China(No.2016YFD0500301,No.2020YFC0842100);the National Major Project for Control and Pre-vention of Infectious Disease in China(No.2018ZX10101002). 摘　　要：With the development of the COVID-19 epidemic,there is an urgent need to establish a system for determining the effectiveness and neutralizing activity of vaccine candidates in biosafety level 2(BSL-2)facilities.Previously,researchers had developed a pseudotyped virus systemfor SARS-CoV andMERS-CoV,based onHIV-1 core,bearing virus spike protein.During the development of a pseudotyped SARS-CoV-2 system,a eukaryotic expression plasmid expressing SARSCoV-2 spike(S)protein was constructed and then co-transfectedwith HIV-1 based plasmid which containing the firefly luciferase reporter gene,into HEK293T cells to prepare the pseudotyped SARS-CoV-2 virus(ppSARS-2).We have successfully established the pseudotyped SARS-CoV-2 system for neutralization and entry inhibition assays.Huh7.5 cell line was found to be the most susceptible to our pseudotyped virus model.Different levels of neutralizing antibodies were detected in convalescent serum samples of COVID-19 patients using ppSARS-2.The recombinant,soluble,angiotensin-converting enzyme 2 protein was found to inhibit the entry of ppSARS-2 in Huh7.5 cells effectively.Furthermore,the neutralization results for ppSARS-2 were consistent with those of live SARS-CoV-2 and determined using the serum samples fromconvalescent patients.In conclusion,we have developed an easily accessible and reliable tool for studying the neutralizing efficiency of antibodies against SARS-CoV-2 and the entry process of the virus in a BSL-2 laboratory. 关键词：Pseudotyped PSEUDOVIRUS SARS-CoV-2 COVID-19 Neutralization assay Viral entry assay

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Development and effectiveness of pseudotyped SARS-CoV-2 system as determined by neutralizing efficiency and entry inhibition test in vitro

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