Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2
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Pseudoviruses are useful virological tools because of their safety and versatility, especially for emerging and re-emerging viruses. Due to its high pathogenicity and infectivity and the lack of effective vaccines and therapeutics, live SARS-CoV-2 has to be handled under biosafety level 3 conditions, which has hindered the development of vaccines and therapeutics. Based on a VSV pseudovirus production system, a pseudovirus-based neutralization assay has been developed for evaluating neutralizing antibodies against SARS-CoV-2 in biosafety level 2 facilities. The key parameters for this assay were optimized, including cell types, cell numbers, virus inoculum. When tested against the SARS-CoV-2 pseudovirus, SARS-CoV-2 convalescent patient sera showed high neutralizing potency, which underscore its potential as therapeutics. The limit of detection for this assay was determined as 22.1 and 43.2 for human and mouse serum samples respectively using a panel of 120 negative samples. The cutoff values were set as 30 and 50 for human and mouse serum samples, respectively. This assay showed relatively low coefficient of variations with 15.9% and 16.2% for the intra- and inter-assay analyses respectively. Taken together, we established a robust pseudovirus-based neutralization assay for SARS-CoV-2 and are glad to share pseudoviruses and related protocols with the developers of vaccines or therapeutics to fight against this lethal virus.
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RBD protein expressed from mRNA functionally inhibited entry of a VSV-based pseudovirus expressing the SARS-CoV-2 S protein in Huh7 cells.
“and functionally inhibited entry of a vesicular stomatitis virus (VSV)-based pseudovirus expressing the SARS-CoV-2 S protein (Nie et al., 2020) in Huh7 cells (Figure 1D).”
From A thermostable mRNA vaccine against COVID-19 (2020) · cites this paper as Nie et al., 2020
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- Authors
- Nie, J., Li, Q., Wu, J., Zhao, C., Hao, H., Liu, H., Zhang, L., Nie, L., Qin, H., Wang, M., Lu, Q., Li, X., Sun, Q., Liu, J., Fan, C., Huang, W., Xu, M., & Wang, Y.
- Affiliations
- National Institutes for Food and Drug Control (CN)
- Chinese Academy of Medical Sciences & Peking Union Medical College (CN)
- Corresponding author
- Huang, Weijin
- Xu, Miao
- Wang, Youchun
- ORCID
- Venue
- Emerging Microbes & Infections
- Publisher
- Informa UK Limited
- Published
- 2020-01
- Volume
- 9
- Issue
- 1
- Pages
- 680-686
- ISSN
- 2222-1751 (ISSN-L)
- PMC
- PMC7144318
- OpenAlex
- W3012650581
- Cited by
- 711
- References
- 27
- Citations by year
- 2026: 21 · 2025: 37 · 2024: 73 · 2023: 95 · 2022: 157 · 2021: 186 · 2020: 141
- FWCI
- 19.06
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Emerging Microbes & Infections (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Informationszentrum Lebenswissenschaften (ZB MED) (submittedVersion, cc-by)
- Topics
- Health Sciences › Medicine › Infectious Diseases › SARS-CoV-2 and COVID-19 Research
- Health Sciences › Medicine › Infectious Diseases › SARS-CoV-2 detection and testing
- Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Virus Infections Studies
- Keywords
- SARS-CoV-2, pseudovirus neutralization assay, neutralizing antibodies, biosafety level 2, convalescent sera, assay validation, detection limit, cut-off values, COVID-19 treatment
- MeSH terms
- Betacoronavirus, COVID-19, SARS-CoV-2, COVID-19 Serotherapy, Animals, Antibodies, Viral, Cell Line, Humans, Immune Sera, Immunization, Passive, Membrane Glycoproteins, Neutralization Tests, Plasmids, Pneumonia, Viral, Sensitivity and Specificity, Vesicular stomatitis Indiana virus, Viral Envelope Proteins, Reproducibility of Results, Coronavirus Infections, Mice, Virus Internalization, Antibodies, Neutralizing, Limit of Detection, Spike Glycoprotein, Coronavirus
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