Blog
Feedback
schliessen

Filtern

 

Bibliotheken

Logo der Bibliothek

Logo FID Pharmazie PubPharm Discovery System Universitätsbibliothek Braunschweig Institut für Informationssysteme

Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2

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

Year of Publication: 2020
Contained in: Emerging microbes & infections Vol. 9, No. 1 (2020), p. 680-686
All journal articles: Search for all articles in this journal
Language: English
Contributors: Nie, Jianhui | Author
Li, Qianqian
Wu, Jiajing
Zhao, Chenyan
Hao, Huan
Liu, Huan
Zhang, Li
Nie, Lingling
Qin, Haiyang
Wang, Meng
Lu, Qiong
Li, Xiaoyu
Sun, Qiyu
Liu, Junkai
Fan, Changfa
Huang, Weijin
Xu, Miao
Wang, Youchun
Full text access:
Electronic availability is being checked...
Links: Full Text (dx.doi.org)
Keywords: COVID-19
G protein, vesicular stomatitis virus
Journal Article
SARS-CoV-2
Validation Study
neutralization assay
neutralizing antibody
pseudovirus
spike protein, SARS-CoV-2
Additional Keywords: *Neutralization Tests
Animals
Antibodies, Neutralizing
Antibodies, Viral
Betacoronavirus
Cell Line
Coronavirus Infections
Humans
Immune Sera
Immunization, Passive
Limit of Detection
Membrane Glycoproteins
Mice
Plasmids
Pneumonia, Viral
Reproducibility of Results
Sensitivity and Specificity
Spike Glycoprotein, Coronavirus
Vesicular stomatitis Indiana virus
Viral Envelope Proteins
Virus Internalization
ISSN: 2222-1751
Note: Copyright: From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Notes: Date Completed 26.03.2020
Date Revised 23.04.2020
published: Print
Citation Status MEDLINE
Copyright: From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Physical Description: Online-Ressource
ID (e.g. DOI, URN): 10.1080/22221751.2020.1743767
more publication details ...

Associated Publications

  • Associated records are being queried...
more (+)
Internes Format
LEADER 05444nma a2201165 c 4500
001 NLM30877650X
003 DE-601
005 20200424215627.0
007 cr uuu---uuuuu
008 200330s2020 000 0 eng d
024 7 |a 10.1080/22221751.2020.1743767  |2 doi 
028 5 2 |a pubmed20n1178.xml 
035 |a (DE-599)NLM32207377 
040 |b ger  |c GBVCP 
041 0 |a eng 
100 1 |a Nie, Jianhui 
245 1 0 |a Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2  |h Elektronische Ressource 
300 |a Online-Ressource 
500 |a Date Completed 26.03.2020 
500 |a Date Revised 23.04.2020 
500 |a published: Print 
500 |a Citation Status MEDLINE 
500 |a Copyright: From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine 
520 |a 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 
611 2 7 |a Journal Article  |2 gnd 
611 2 7 |a Validation Study  |2 gnd 
653 2 |a Animals  |6 D000818 
653 2 |a Antibodies, Neutralizing  |6 D057134  |a *immunology  |6 Q000276 
653 2 |a Antibodies, Viral  |6 D000914  |a *immunology  |6 Q000276 
653 2 |a Betacoronavirus  |6 D000073640  |a *immunology  |6 Q000276 
653 2 |a Cell Line  |6 D002460 
653 2 |a Coronavirus Infections  |6 D018352  |a *immunology  |6 Q000276  |a therapy  |6 Q000628 
653 2 |a Humans  |6 D006801 
653 2 |a Immune Sera  |6 D007106  |a *immunology  |6 Q000276 
653 2 |a Immunization, Passive  |6 D007116 
653 2 |a Limit of Detection  |6 D057230 
653 2 |a Membrane Glycoproteins  |6 D008562  |a immunology  |6 Q000276 
653 2 |a Mice  |6 D051379 
653 2 |a *Neutralization Tests  |6 D009500 
653 2 |a Plasmids  |6 D010957 
653 2 |a Pneumonia, Viral  |6 D011024  |a *immunology  |6 Q000276 
653 2 |a Reproducibility of Results  |6 D015203 
653 2 |a Sensitivity and Specificity  |6 D012680 
653 2 |a Spike Glycoprotein, Coronavirus  |6 D064370  |a genetics  |6 Q000235  |a *immunology  |6 Q000276 
653 2 |a Vesicular stomatitis Indiana virus  |6 D014721  |a genetics  |6 Q000235 
653 2 |a Viral Envelope Proteins  |6 D014759  |a immunology  |6 Q000276 
653 2 |a Virus Internalization  |6 D053586 
655 7 |a COVID-19  |2 gnd 
655 7 |a SARS-CoV-2  |2 gnd 
655 7 |a neutralization assay  |2 gnd 
655 7 |a neutralizing antibody  |2 gnd 
655 7 |a pseudovirus  |2 gnd 
655 7 |a Antibodies, Neutralizing  |2 gnd 
655 7 |a Antibodies, Viral  |2 gnd 
655 7 |a G protein, vesicular stomatitis virus  |2 gnd 
655 7 |a Immune Sera  |2 gnd 
655 7 |a Membrane Glycoproteins  |2 gnd 
655 7 |a Spike Glycoprotein, Coronavirus  |2 gnd 
655 7 |a Viral Envelope Proteins  |2 gnd 
655 7 |a spike protein, SARS-CoV-2  |2 gnd 
689 0 0 |A f  |a Journal Article 
689 0 1 |A f  |a Validation Study 
689 0 |5 DE-601 
689 1 0 |a COVID-19 
689 1 1 |a SARS-CoV-2 
689 1 2 |a neutralization assay 
689 1 3 |a neutralizing antibody 
689 1 4 |a pseudovirus 
689 1 |5 DE-601 
689 2 0 |a Antibodies, Neutralizing 
689 2 1 |a Antibodies, Viral 
689 2 2 |a G protein, vesicular stomatitis virus 
689 2 3 |a Immune Sera 
689 2 4 |a Membrane Glycoproteins 
689 2 5 |a Spike Glycoprotein, Coronavirus 
689 2 6 |a Viral Envelope Proteins 
689 2 7 |a spike protein, SARS-CoV-2 
689 2 |5 DE-601 
700 1 |a Li, Qianqian 
700 1 |a Wu, Jiajing 
700 1 |a Zhao, Chenyan 
700 1 |a Hao, Huan 
700 1 |a Liu, Huan 
700 1 |a Zhang, Li 
700 1 |a Nie, Lingling 
700 1 |a Qin, Haiyang 
700 1 |a Wang, Meng 
700 1 |a Lu, Qiong 
700 1 |a Li, Xiaoyu 
700 1 |a Sun, Qiyu 
700 1 |a Liu, Junkai 
700 1 |a Fan, Changfa 
700 1 |a Huang, Weijin 
700 1 |a Xu, Miao 
700 1 |a Wang, Youchun 
773 0 8 |i in  |t Emerging microbes & infections  |g Vol. 9, No. 1 (2020), p. 680-686  |q 9:1<680-686  |w (DE-601)NLM249572478  |x 2222-1751 
856 4 1 |u http://dx.doi.org/10.1080/22221751.2020.1743767  |3 Volltext 
912 |a GBV_NLM 
951 |a AR 
952 |d 9  |j 2020  |e 1  |b 23  |c 12  |h 680-686