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Evaluation of Cell-Based and Surrogate SARS-CoV-2 Neutralization Assays
- Source :
- Journal of Clinical Microbiology
- Publication Year :
- 2021
-
Abstract
- Determinants of protective immunity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection require the development of well-standardized, reproducible antibody assays. This need has led to the emergence of a variety of neutralization assays. Head-to-head evaluation of different SARS-CoV-2 neutralization platforms could facilitate comparisons across studies and laboratories. Five neutralization assays were compared using 40 plasma samples from convalescent individuals with mild to moderate coronavirus disease 2019 (COVID-19): four cell-based systems using either live recombinant SARS-CoV-2 or pseudotyped viral particles created with lentivirus (LV) or vesicular stomatitis virus (VSV) packaging and one surrogate enzyme-linked immunosorbent assay (ELISA)-based test that measures inhibition of the spike protein receptor binding domain (RBD) binding its receptor human angiotensin converting enzyme 2 (hACE2). Vero cells, Vero E6 cells, HEK293T cells expressing hACE2, and TZM-bl cells expressing hACE2 and transmembrane serine protease 2 were tested. All cell-based assays showed 50% neutralizing dilution (ND50) geometric mean titers (GMTs) that were highly correlated (Pearson r = 0.81 to 0.89) and ranged within 3.4-fold. The live virus assay and LV pseudovirus assays with HEK293T/hACE2 cells showed very similar mean titers, 141 and 178, respectively. ND50 titers positively correlated with plasma IgG targeting SARS-CoV-2 spike protein and RBD (r = 0.63 to 0.89), but moderately correlated with nucleoprotein IgG (r = 0.46 to 0.73). ND80 GMTs mirrored ND50 data and showed similar correlation between assays and with IgG concentrations. The VSV pseudovirus assay and LV pseudovirus assay with HEK293T/hACE2 cells in low- and high-throughput versions were calibrated against the WHO SARS-CoV-2 IgG standard. High concordance between the outcomes of cell-based assays with live and pseudotyped virions enables valid cross-study comparison using these platforms.
- Subjects :
- Microbiology (medical)
viruses
Antibodies, Viral
Neutralization
law.invention
law
Neutralization Tests
neutralization assay
Virology
antibody
Chlorocebus aethiops
Animals
Humans
Vero Cells
biology
Chemistry
SARS-CoV-2
COVID-19
biology.organism_classification
Antibodies, Neutralizing
Nucleoprotein
Titer
HEK293 Cells
Vesicular stomatitis virus
Lentivirus
Spike Glycoprotein, Coronavirus
Vero cell
Recombinant DNA
biology.protein
Antibody
Subjects
Details
- ISSN :
- 1098660X
- Volume :
- 59
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- Journal of clinical microbiology
- Accession number :
- edsair.doi.dedup.....32f66dbd89812d4e1725fc2b6b6e800f