Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells

Xiong, Hua-Long1,2; Wu, Yang-Tao1,2; Cao, Jia-Li1,2; Yang, Ren3; Liu, Ying-Xia4; Ma, Jian1,2; Qiao, Xiao-Yang1,2; Yao, Xiang-Yang5; Zhang, Bao-Hui1,2; Zhang, Ya-Li1,2; Hou, Wang-Heng1,2; Shi, Yang1,2; Xu, Jing-Jing6; Zhang, Liang1,2; Wang, Shao-Juan1,2; Fu, Bao-Rong1,2; Yang, Ting6; Ge, Sheng-Xiang1,2; Zhang, Jun1,2; Yuan, Quan1,2; Huang, Bao-Ying3; Li, Zhi-Yong5; Zhang, Tian-Ying1,2; Xia, Ning-Shao1,2

2020-01

10.1080/22221751.2020.1815589

Emerging Microbes & Infections   影响因子和分区

2222-1751

The global pandemic of coronavirus disease 2019 (COVID-19) is a disaster for human society. A convenient and reliable neutralization assay is very important for the development of vaccines and novel drugs. In this study, a G protein-deficient vesicular stomatitis virus (VSVdG) bearing a truncated spike protein (S with C-terminal 18 amino acid truncation) was compared to that bearing the full-length spike protein of SARS-CoV-2 and showed much higher efficiency. A neutralization assay was established based on VSV-SARS-CoV-2-Sdel18 pseudovirus and hACE2-overexpressing BHK21 cells (BHK21-hACE2 cells). The experimental results can be obtained by automatically counting the number of EGFP-positive cells at 12 h after infection, making the assay convenient and high-throughput. The serum neutralizing titer measured by the VSV-SARS-CoV-2-Sdel18 pseudovirus assay has a good correlation with that measured by the wild type SARS-CoV-2 assay. Seven neutralizing monoclonal antibodies targeting the receptor binding domain (RBD) of the SARS-CoV-2 S protein were obtained. This efficient and reliable pseudovirus assay model could facilitate the development of new drugs and vaccines.

SARS-CoV-2 pseudovirus neutralization assay vesicular stomatitis virus VSV-SARS-CoV-2

SCI

英语

其他

National Natural Science Foundation of China[81993149041][81702006] ; Science and Technology Major Project of the Fujian Province[2020YZ014001] ; Xiamen Science and Technology Major Project[3502Z2020YJ02] ; National Key Research and Development Program of China[2016YFD0500301]

Immunology ; Microbiology

Immunology ; Microbiology

WOS:000576054800001

TAYLOR & FRANCIS LTD

Web of Science

1.Xiamen Univ, Sch Publ Hlth, Natl Inst Diagnost & Vaccine Dev Infect Dis, State Key Lab Mol Vaccinol & Mol Diagnost, Xiamen 361102, Peoples R China
2.Xiamen Univ, Sch Life Sci, Xiamen 361102, Peoples R China
3.China Cdc, NHC Key Lab Biosafety, Natl Inst Viral Dis Control & Prevent, Beijing 102206, Peoples R China
4.Southern Univ Sci & Technol, State Key Discipline Infect Dis, Shenzhen Peoples Hosp 3, Hosp 2,Natl Clin Res Ctr Infect Dis,Shenzhen Key, Shenzhen, Peoples R China
5.Xiamen Univ, Hosp 1, Xiamen 361003, Peoples R China
6.Fujian Med Univ, Dept Hematol, Fujian Inst Hematol, Union Hosp,Fujian Prov Key Lab Hematol, Fuzhou, Peoples R China

Xiong, Hua-Long,Wu, Yang-Tao,Cao, Jia-Li,et al. Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells[J]. Emerging Microbes & Infections,2020,9(1):2105-2113.

Xiong, Hua-Long.,Wu, Yang-Tao.,Cao, Jia-Li.,Yang, Ren.,Liu, Ying-Xia.,...&Xia, Ning-Shao.(2020).Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells.Emerging Microbes & Infections,9(1),2105-2113.

Xiong, Hua-Long,et al."Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells".Emerging Microbes & Infections 9.1(2020):2105-2113.