Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2
Identifieur interne : 000663 ( Main/Exploration ); précédent : 000662; suivant : 000664Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2
Auteurs : Jianhui Nie [République populaire de Chine] ; Qianqian Li [République populaire de Chine] ; Jiajing Wu [République populaire de Chine] ; Chenyan Zhao [République populaire de Chine] ; Huan Hao [République populaire de Chine] ; Huan Liu [République populaire de Chine] ; Li Zhang [République populaire de Chine] ; Lingling Nie [République populaire de Chine] ; Haiyang Qin [République populaire de Chine] ; Meng Wang [République populaire de Chine] ; Qiong Lu [République populaire de Chine] ; Xiaoyu Li [République populaire de Chine] ; Qiyu Sun [République populaire de Chine] ; Junkai Liu [République populaire de Chine] ; Changfa Fan [République populaire de Chine] ; Weijin Huang [République populaire de Chine] ; Miao Xu [République populaire de Chine] ; Youchun Wang [République populaire de Chine]Source :
- Emerging Microbes & Infections [ 2222-1751 ] ; 2020.
Descripteurs français
- KwdFr :
- Animaux, Anticorps antiviraux (immunologie), Anticorps neutralisants (immunologie), Glycoprotéine de spicule des coronavirus (génétique), Glycoprotéine de spicule des coronavirus (immunologie), Glycoprotéines membranaires (immunologie), Humains, Immunisation passive, Infections à coronavirus (), Infections à coronavirus (immunologie), Lignée cellulaire, Limite de détection, Plasmides, Pneumopathie virale (immunologie), Protéines de l'enveloppe virale (immunologie), Pénétration virale, Reproductibilité des résultats, Sensibilité et spécificité, Souris, Sérums immuns (immunologie), Tests de neutralisation, Virus de la stomatite vésiculeuse de type Indiana (génétique).
- MESH :
- génétique : Glycoprotéine de spicule des coronavirus, Virus de la stomatite vésiculeuse de type Indiana.
- immunologie : Anticorps antiviraux, Anticorps neutralisants, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires, Infections à coronavirus, Pneumopathie virale, Protéines de l'enveloppe virale, Sérums immuns.
- Animaux, Humains, Immunisation passive, Infections à coronavirus, Lignée cellulaire, Limite de détection, Plasmides, Pénétration virale, Reproductibilité des résultats, Sensibilité et spécificité, Souris, Tests de neutralisation.
English descriptors
- KwdEn :
- Animals, Antibodies, Neutralizing (immunology), Antibodies, Viral (immunology), Betacoronavirus (immunology), Cell Line, Coronavirus Infections (immunology), Coronavirus Infections (therapy), Humans, Immune Sera (immunology), Immunization, Passive, Limit of Detection, Membrane Glycoproteins (immunology), Mice, Neutralization Tests, Plasmids, Pneumonia, Viral (immunology), Reproducibility of Results, Sensitivity and Specificity, Spike Glycoprotein, Coronavirus (genetics), Spike Glycoprotein, Coronavirus (immunology), Vesicular stomatitis Indiana virus (genetics), Viral Envelope Proteins (immunology), Virus Internalization.
- MESH :
- chemical , genetics : Spike Glycoprotein, Coronavirus.
- chemical , immunology : Antibodies, Neutralizing, Antibodies, Viral, Immune Sera, Membrane Glycoproteins, Spike Glycoprotein, Coronavirus, Viral Envelope Proteins.
- genetics : Vesicular stomatitis Indiana virus.
- immunology : Betacoronavirus, Coronavirus Infections, Pneumonia, Viral.
- therapy : Coronavirus Infections.
- Animals, Cell Line, Humans, Immunization, Passive, Limit of Detection, Mice, Neutralization Tests, Plasmids, Reproducibility of Results, Sensitivity and Specificity, Virus Internalization.
Abstract
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.
Url:
DOI: 10.1080/22221751.2020.1743767
PubMed: 32207377
PubMed Central: 7144318
Affiliations:
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China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="AF3"><institution>Wuhan Institute of Biological Products</institution>, Wuhan,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Zhao, Chenyan" sort="Zhao, Chenyan" uniqKey="Zhao C" first="Chenyan" last="Zhao">Chenyan Zhao</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Hao, Huan" sort="Hao, Huan" uniqKey="Hao H" first="Huan" last="Hao">Huan Hao</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Liu, Huan" sort="Liu, Huan" uniqKey="Liu H" first="Huan" last="Liu">Huan Liu</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Zhang, Li" sort="Zhang, Li" uniqKey="Zhang L" first="Li" last="Zhang">Li Zhang</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Nie, Lingling" sort="Nie, Lingling" uniqKey="Nie L" first="Lingling" last="Nie">Lingling Nie</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Qin, Haiyang" sort="Qin, Haiyang" uniqKey="Qin H" first="Haiyang" last="Qin">Haiyang Qin</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Wang, Meng" sort="Wang, Meng" uniqKey="Wang M" first="Meng" last="Wang">Meng Wang</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Lu, Qiong" sort="Lu, Qiong" uniqKey="Lu Q" first="Qiong" last="Lu">Qiong Lu</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Li, Xiaoyu" sort="Li, Xiaoyu" uniqKey="Li X" first="Xiaoyu" last="Li">Xiaoyu Li</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Sun, Qiyu" sort="Sun, Qiyu" uniqKey="Sun Q" first="Qiyu" last="Sun">Qiyu Sun</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Liu, Junkai" sort="Liu, Junkai" uniqKey="Liu J" first="Junkai" last="Liu">Junkai Liu</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Fan, Changfa" sort="Fan, Changfa" uniqKey="Fan C" first="Changfa" last="Fan">Changfa Fan</name><affiliation wicri:level="1"><nlm:aff id="AF4"><institution>Division of Animal Model Research, Institute for Laboratory Animal Resources, National Institutes for Food and Drug Control</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Huang, Weijin" sort="Huang, Weijin" uniqKey="Huang W" first="Weijin" last="Huang">Weijin Huang</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Xu, Miao" sort="Xu, Miao" uniqKey="Xu M" first="Miao" last="Xu">Miao Xu</name><affiliation wicri:level="1"><nlm:aff id="AF5"><institution>Institute for Biological Product Control, National Institutes for Food and Drug Control</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Wang, Youchun" sort="Wang, Youchun" uniqKey="Wang Y" first="Youchun" last="Wang">Youchun Wang</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="AF2"><institution>Graduate School of Peking Union Medical College</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de 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Nie</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Li, Qianqian" sort="Li, Qianqian" uniqKey="Li Q" first="Qianqian" last="Li">Qianqian Li</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="AF2"><institution>Graduate School of Peking Union Medical College</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Wu, Jiajing" sort="Wu, Jiajing" uniqKey="Wu J" first="Jiajing" last="Wu">Jiajing Wu</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="AF3"><institution>Wuhan Institute of Biological Products</institution>, Wuhan,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Zhao, Chenyan" sort="Zhao, Chenyan" uniqKey="Zhao C" first="Chenyan" last="Zhao">Chenyan Zhao</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Hao, Huan" sort="Hao, Huan" uniqKey="Hao H" first="Huan" last="Hao">Huan Hao</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Liu, Huan" sort="Liu, Huan" uniqKey="Liu H" first="Huan" last="Liu">Huan Liu</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Zhang, Li" sort="Zhang, Li" uniqKey="Zhang L" first="Li" last="Zhang">Li Zhang</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Nie, Lingling" sort="Nie, Lingling" uniqKey="Nie L" first="Lingling" last="Nie">Lingling Nie</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Qin, Haiyang" sort="Qin, Haiyang" uniqKey="Qin H" first="Haiyang" last="Qin">Haiyang Qin</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Wang, Meng" sort="Wang, Meng" uniqKey="Wang M" first="Meng" last="Wang">Meng Wang</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Lu, Qiong" sort="Lu, Qiong" uniqKey="Lu Q" first="Qiong" last="Lu">Qiong Lu</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Li, Xiaoyu" sort="Li, Xiaoyu" uniqKey="Li X" first="Xiaoyu" last="Li">Xiaoyu Li</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Sun, Qiyu" sort="Sun, Qiyu" uniqKey="Sun Q" first="Qiyu" last="Sun">Qiyu Sun</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Liu, Junkai" sort="Liu, Junkai" uniqKey="Liu J" first="Junkai" last="Liu">Junkai Liu</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Fan, Changfa" sort="Fan, Changfa" uniqKey="Fan C" first="Changfa" last="Fan">Changfa Fan</name><affiliation wicri:level="1"><nlm:aff id="AF4"><institution>Division of Animal Model Research, Institute for Laboratory Animal Resources, National Institutes for Food and Drug Control</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Huang, Weijin" sort="Huang, Weijin" uniqKey="Huang W" first="Weijin" last="Huang">Weijin Huang</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Xu, Miao" sort="Xu, Miao" uniqKey="Xu M" first="Miao" last="Xu">Miao Xu</name><affiliation wicri:level="1"><nlm:aff id="AF5"><institution>Institute for Biological Product Control, National Institutes for Food and Drug Control</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Wang, Youchun" sort="Wang, Youchun" uniqKey="Wang Y" first="Youchun" last="Wang">Youchun Wang</name><affiliation wicri:level="1"><nlm:aff id="AF1"><institution>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC)</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="AF2"><institution>Graduate School of Peking Union Medical College</institution>, Beijing,<country>People’s Republic of China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></analytic><series><title level="j">Emerging Microbes & Infections</title><idno type="eISSN">2222-1751</idno><imprint><date when="2020">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Neutralizing (immunology)</term><term>Antibodies, Viral (immunology)</term><term>Betacoronavirus (immunology)</term><term>Cell Line</term><term>Coronavirus Infections (immunology)</term><term>Coronavirus Infections (therapy)</term><term>Humans</term><term>Immune Sera (immunology)</term><term>Immunization, Passive</term><term>Limit of Detection</term><term>Membrane Glycoproteins (immunology)</term><term>Mice</term><term>Neutralization Tests</term><term>Plasmids</term><term>Pneumonia, Viral (immunology)</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term><term>Spike Glycoprotein, Coronavirus (genetics)</term><term>Spike Glycoprotein, Coronavirus (immunology)</term><term>Vesicular stomatitis Indiana virus (genetics)</term><term>Viral Envelope Proteins (immunology)</term><term>Virus Internalization</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Anticorps antiviraux (immunologie)</term><term>Anticorps neutralisants (immunologie)</term><term>Glycoprotéine de spicule des coronavirus (génétique)</term><term>Glycoprotéine de spicule des coronavirus (immunologie)</term><term>Glycoprotéines membranaires (immunologie)</term><term>Humains</term><term>Immunisation passive</term><term>Infections à coronavirus ()</term><term>Infections à coronavirus (immunologie)</term><term>Lignée cellulaire</term><term>Limite de détection</term><term>Plasmides</term><term>Pneumopathie virale (immunologie)</term><term>Protéines de l'enveloppe virale (immunologie)</term><term>Pénétration virale</term><term>Reproductibilité des résultats</term><term>Sensibilité et spécificité</term><term>Souris</term><term>Sérums immuns (immunologie)</term><term>Tests de neutralisation</term><term>Virus de la stomatite vésiculeuse de type Indiana (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Spike Glycoprotein, Coronavirus</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Neutralizing</term><term>Antibodies, Viral</term><term>Immune Sera</term><term>Membrane Glycoproteins</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Vesicular stomatitis Indiana virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Glycoprotéine de spicule des coronavirus</term><term>Virus de la stomatite vésiculeuse de type Indiana</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiviraux</term><term>Anticorps neutralisants</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires</term><term>Infections à coronavirus</term><term>Pneumopathie virale</term><term>Protéines de l'enveloppe virale</term><term>Sérums immuns</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Betacoronavirus</term><term>Coronavirus Infections</term><term>Pneumonia, Viral</term></keywords><keywords scheme="MESH" qualifier="therapy" xml:lang="en"><term>Coronavirus Infections</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cell Line</term><term>Humans</term><term>Immunization, Passive</term><term>Limit of Detection</term><term>Mice</term><term>Neutralization Tests</term><term>Plasmids</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term><term>Virus Internalization</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Humains</term><term>Immunisation passive</term><term>Infections à coronavirus</term><term>Lignée cellulaire</term><term>Limite de détection</term><term>Plasmides</term><term>Pénétration virale</term><term>Reproductibilité des résultats</term><term>Sensibilité et spécificité</term><term>Souris</term><term>Tests de neutralisation</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title>ABSTRACT</title><p>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.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Hui Ds, Eia" uniqKey="Hui Ds E">EIA Hui DS</name></author><author><name sortKey="Madani, Ta" uniqKey="Madani T">TA Madani</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Li, Q" uniqKey="Li Q">Q Li</name></author><author><name sortKey="Guan, X" uniqKey="Guan X">X Guan</name></author><author><name sortKey="Wu, P" uniqKey="Wu P">P Wu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Phelan, Al" uniqKey="Phelan A">AL Phelan</name></author><author><name sortKey="Katz, R" uniqKey="Katz R">R Katz</name></author><author><name sortKey="Gostin, Lo" uniqKey="Gostin L">LO. 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