Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2.
Identifieur interne : 000781 ( an2020/Analysis ); précédent : 000780; suivant : 000782Establishment 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.
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.
DOI: 10.1080/22221751.2020.1743767
PubMed: 32207377
Affiliations:
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first="Weijin" last="Huang">Weijin Huang</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Xu, Miao" sort="Xu, Miao" uniqKey="Xu M" first="Miao" last="Xu">Miao Xu</name><affiliation wicri:level="3"><nlm:affiliation>Institute for Biological Product Control, National Institutes for Food and Drug Control, Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de 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Q" first="Qianqian" last="Li">Qianqian Li</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Wu, Jiajing" sort="Wu, Jiajing" uniqKey="Wu J" first="Jiajing" last="Wu">Jiajing Wu</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Zhao, Chenyan" sort="Zhao, Chenyan" uniqKey="Zhao C" first="Chenyan" last="Zhao">Chenyan Zhao</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Hao, Huan" sort="Hao, Huan" uniqKey="Hao H" first="Huan" last="Hao">Huan Hao</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Liu, Huan" sort="Liu, Huan" uniqKey="Liu H" first="Huan" last="Liu">Huan Liu</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Zhang, Li" sort="Zhang, Li" uniqKey="Zhang L" first="Li" last="Zhang">Li Zhang</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Nie, Lingling" sort="Nie, Lingling" uniqKey="Nie L" first="Lingling" last="Nie">Lingling Nie</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Qin, Haiyang" sort="Qin, Haiyang" uniqKey="Qin H" first="Haiyang" last="Qin">Haiyang Qin</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Wang, Meng" sort="Wang, Meng" uniqKey="Wang M" first="Meng" last="Wang">Meng Wang</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Lu, Qiong" sort="Lu, Qiong" uniqKey="Lu Q" first="Qiong" last="Lu">Qiong Lu</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Li, Xiaoyu" sort="Li, Xiaoyu" uniqKey="Li X" first="Xiaoyu" last="Li">Xiaoyu Li</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Sun, Qiyu" sort="Sun, Qiyu" uniqKey="Sun Q" first="Qiyu" last="Sun">Qiyu Sun</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Liu, Junkai" sort="Liu, Junkai" uniqKey="Liu J" first="Junkai" last="Liu">Junkai Liu</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Fan, Changfa" sort="Fan, Changfa" uniqKey="Fan C" first="Changfa" last="Fan">Changfa Fan</name><affiliation wicri:level="3"><nlm:affiliation>Division of Animal Model Research, Institute for Laboratory Animal Resources, National Institutes for Food and Drug Control, Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of Animal Model Research, Institute for Laboratory Animal Resources, National Institutes for Food and Drug Control, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Huang, Weijin" sort="Huang, Weijin" uniqKey="Huang W" first="Weijin" last="Huang">Weijin Huang</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Xu, Miao" sort="Xu, Miao" uniqKey="Xu M" first="Miao" last="Xu">Miao Xu</name><affiliation wicri:level="3"><nlm:affiliation>Institute for Biological Product Control, National Institutes for Food and Drug Control, Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute for Biological Product Control, National Institutes for Food and Drug Control, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Wang, Youchun" sort="Wang, Youchun" uniqKey="Wang Y" first="Youchun" last="Wang">Youchun Wang</name><affiliation wicri:level="3"><nlm:affiliation>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author></analytic><series><title level="j">Emerging microbes & infections</title><idno type="eISSN">2222-1751</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Nie, Jianhui" sort="Nie, Jianhui" uniqKey="Nie J" first="Jianhui" last="Nie">Jianhui Nie</name></noRegion><name sortKey="Fan, Changfa" sort="Fan, Changfa" uniqKey="Fan C" first="Changfa" last="Fan">Changfa Fan</name><name sortKey="Hao, Huan" sort="Hao, Huan" uniqKey="Hao H" first="Huan" last="Hao">Huan Hao</name><name sortKey="Huang, Weijin" sort="Huang, Weijin" uniqKey="Huang W" first="Weijin" last="Huang">Weijin Huang</name><name sortKey="Li, Qianqian" sort="Li, Qianqian" uniqKey="Li Q" first="Qianqian" last="Li">Qianqian Li</name><name sortKey="Li, Xiaoyu" sort="Li, Xiaoyu" uniqKey="Li X" first="Xiaoyu" last="Li">Xiaoyu Li</name><name sortKey="Liu, Huan" sort="Liu, Huan" uniqKey="Liu H" first="Huan" last="Liu">Huan Liu</name><name sortKey="Liu, Junkai" sort="Liu, Junkai" uniqKey="Liu J" first="Junkai" last="Liu">Junkai Liu</name><name sortKey="Lu, Qiong" sort="Lu, Qiong" uniqKey="Lu Q" first="Qiong" last="Lu">Qiong Lu</name><name sortKey="Nie, Lingling" sort="Nie, Lingling" uniqKey="Nie L" first="Lingling" last="Nie">Lingling Nie</name><name sortKey="Qin, Haiyang" sort="Qin, Haiyang" uniqKey="Qin H" first="Haiyang" last="Qin">Haiyang Qin</name><name sortKey="Sun, Qiyu" sort="Sun, Qiyu" uniqKey="Sun Q" first="Qiyu" last="Sun">Qiyu Sun</name><name sortKey="Wang, Meng" sort="Wang, Meng" uniqKey="Wang M" first="Meng" last="Wang">Meng Wang</name><name sortKey="Wang, Youchun" sort="Wang, Youchun" uniqKey="Wang Y" first="Youchun" last="Wang">Youchun Wang</name><name sortKey="Wu, Jiajing" sort="Wu, Jiajing" uniqKey="Wu J" first="Jiajing" last="Wu">Jiajing Wu</name><name sortKey="Xu, Miao" sort="Xu, Miao" uniqKey="Xu M" first="Miao" last="Xu">Miao Xu</name><name sortKey="Zhang, Li" sort="Zhang, Li" uniqKey="Zhang L" first="Li" last="Zhang">Li Zhang</name><name sortKey="Zhao, Chenyan" sort="Zhao, Chenyan" uniqKey="Zhao C" first="Chenyan" last="Zhao">Chenyan Zhao</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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