A Novel In-Cell ELISA Assay Allows Rapid and Automated Quantification of SARS-CoV-2 to Analyze Neutralizing Antibodies and Antiviral Compounds.
Animals
Antibodies, Neutralizing
/ blood
Antibodies, Viral
/ blood
Antiviral Agents
/ pharmacology
COVID-19
/ diagnosis
COVID-19 Testing
/ methods
Caco-2 Cells
Chlorocebus aethiops
Diagnostic Tests, Routine
/ methods
Enzyme-Linked Immunosorbent Assay
/ methods
Humans
Immunization, Passive
Neutralization Tests
/ methods
SARS-CoV-2
/ genetics
Vero Cells
Virus Replication
/ drug effects
COVID-19 Serotherapy
COVID-19
SARS-CoV-2
antibodies
interferon
neutralizing
vaccine
Journal
Frontiers in immunology
ISSN: 1664-3224
Titre abrégé: Front Immunol
Pays: Switzerland
ID NLM: 101560960
Informations de publication
Date de publication:
2020
2020
Historique:
received:
23
07
2020
accepted:
24
09
2020
entrez:
9
11
2020
pubmed:
10
11
2020
medline:
15
12
2020
Statut:
epublish
Résumé
The coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is currently the most pressing medical and socioeconomic challenge. Constituting important correlates of protection, the determination of virus-neutralizing antibodies (NAbs) is indispensable for convalescent plasma selection, vaccine candidate evaluation, and immunity certificates. In contrast to standard serological ELISAs, plaque reduction neutralization tests (PRNTs) are laborious, time-consuming, expensive, and restricted to specialized laboratories. To replace microscopic counting-based SARS-CoV-2 PRNTs by a novel assay exempt from genetically modified viruses, which are inapplicable in most diagnostics departments, we established a simple, rapid, and automated SARS-CoV-2 neutralization assay employing an in-cell ELISA (icELISA) approach. After optimization of various parameters such as virus-specific antibodies, cell lines, virus doses, and duration of infection, SARS-CoV-2-infected cells became amenable as direct antigen source for quantitative icELISA. Antiviral agents such as human sera containing NAbs or antiviral interferons dose dependently reduced the SARS-CoV-2-specific signal. Applying increased infectious doses, the icELISA-based neutralization test (icNT) was superior to PRNT in discriminating convalescent sera with high from those with intermediate neutralizing capacities. In addition, the icNT was found to be specific, discriminating between SARS-CoV-2-specific NAbs and those raised against other coronaviruses. Altogether, the SARS-CoV-2 icELISA test allows rapid (<48 h in total, read-out in seconds) and automated quantification of virus infection in cell culture to evaluate the efficacy of NAbs and antiviral drugs using reagents and equipment present in most routine diagnostics departments.
Identifiants
pubmed: 33162987
doi: 10.3389/fimmu.2020.573526
pmc: PMC7581787
doi:
Substances chimiques
Antibodies, Neutralizing
0
Antibodies, Viral
0
Antiviral Agents
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
573526Informations de copyright
Copyright © 2020 Schöler, Le-Trilling, Eilbrecht, Mennerich, Anastasiou, Krawczyk, Herrmann, Dittmer and Trilling.
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