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.

Media Type:

Electronic Article

Year of Publication:

2020

Contained In:

Emerging microbes & infections - Vol. 9, No. 1 (2020), p. 680-686

Language:

English

Contributors:

Nie, Jianhui
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

Links:

Volltext

Keywords:

*Neutralization Tests
Animals
Antibodies, Neutralizing
Antibodies, Viral
Betacoronavirus
COVID-19
Cell Line
Coronavirus Infections
G protein, vesicular stomatitis virus
Humans
Immune Sera
Immunization, Passive
Journal Article
Limit of Detection
Membrane Glycoproteins
Mice
Neutralization assay
Neutralizing antibody
Plasmids
Pneumonia, Viral
Pseudovirus
Reproducibility of Results
SARS-CoV-2
Sensitivity and Specificity
Spike Glycoprotein, Coronavirus
Spike protein, SARS-CoV-2
Validation Study
Vesicular stomatitis Indiana virus
Viral Envelope Proteins
Virus Internalization

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

doi:

10.1080/22221751.2020.1743767

PMID:

32207377

PPN (Catalogue-ID):

NLM30877650X