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Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells.
Emerg Microbes Infect. 2020 Dec; 9(1):2105-2113.EM

Abstract

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.

Authors+Show Affiliations

State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, People's Republic of China.Shenzhen Key Laboratory of Pathogen and Immunity, National Clinical Research Center for Infectious Disease, State Key Discipline of Infectious Disease, Shenzhen Third People's Hospital, Second Hospital Affiliated to Southern University of Science and Technology, Shenzhen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.The First Hospital of Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.Department of Hematology, Fujian Medical University Union Hospital, Fujian Provincial Key Laboratory on Hematology, Fujian Institute of Hematology, Fuzhou, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.Department of Hematology, Fujian Medical University Union Hospital, Fujian Provincial Key Laboratory on Hematology, Fujian Institute of Hematology, Fuzhou, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, People's Republic of China.The First Hospital of Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32893735

Citation

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, vol. 9, no. 1, 2020, pp. 2105-2113.
Xiong HL, Wu YT, Cao JL, et al. Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells. Emerg Microbes Infect. 2020;9(1):2105-2113.
Xiong, H. L., Wu, Y. T., Cao, J. L., Yang, R., Liu, Y. X., Ma, J., Qiao, X. Y., Yao, X. Y., Zhang, B. H., Zhang, Y. L., Hou, W. H., Shi, Y., Xu, J. J., Zhang, L., Wang, S. J., Fu, B. R., Yang, T., Ge, S. X., Zhang, J., ... Xia, N. S. (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. https://doi.org/10.1080/22221751.2020.1815589
Xiong HL, et al. Robust Neutralization Assay Based On SARS-CoV-2 S-protein-bearing Vesicular Stomatitis Virus (VSV) Pseudovirus and ACE2-overexpressing BHK21 Cells. Emerg Microbes Infect. 2020;9(1):2105-2113. PubMed PMID: 32893735.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells. AU - Xiong,Hua-Long, AU - Wu,Yang-Tao, AU - Cao,Jia-Li, AU - Yang,Ren, AU - Liu,Ying-Xia, AU - Ma,Jian, AU - Qiao,Xiao-Yang, AU - Yao,Xiang-Yang, AU - Zhang,Bao-Hui, AU - Zhang,Ya-Li, AU - Hou,Wang-Heng, AU - Shi,Yang, AU - Xu,Jing-Jing, AU - Zhang,Liang, AU - Wang,Shao-Juan, AU - Fu,Bao-Rong, AU - Yang,Ting, AU - Ge,Sheng-Xiang, AU - Zhang,Jun, AU - Yuan,Quan, AU - Huang,Bao-Ying, AU - Li,Zhi-Yong, AU - Zhang,Tian-Ying, AU - Xia,Ning-Shao, PY - 2020/9/8/pubmed PY - 2020/10/6/medline PY - 2020/9/7/entrez KW - SARS-CoV-2 KW - VSV-SARS-CoV-2 KW - neutralization assay KW - pseudovirus KW - vesicular stomatitis virus SP - 2105 EP - 2113 JF - Emerging microbes & infections JO - Emerg Microbes Infect VL - 9 IS - 1 N2 - 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. SN - 2222-1751 UR - https://www.unboundmedicine.com/medline/citation/32893735/Robust_neutralization_assay_based_on_SARS_CoV_2_S_protein_bearing_vesicular_stomatitis_virus__VSV__pseudovirus_and_ACE2_overexpressing_BHK21_cells_ L2 - https://www.tandfonline.com/doi/full/10.1080/22221751.2020.1815589 DB - PRIME DP - Unbound Medicine ER -