Journal
MICROBIOLOGY SPECTRUM
Volume 9, Issue 2, Pages -Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/Spectrum.01059-21
Keywords
SARS-CoV-2; immunoassays; neutralizing antibodies
Categories
Funding
- ALSAC
- National Institute of Allergy and Infectious Diseases under HHS [HHSN27220140006C]
- University of Tennessee Research Foundation
- American Society of Hematology Scholar Award
- NIAID Collaborative Influenza Vaccine Innovation Centers (CIVIC) [75N93019C00051]
- NIAID Center of Excellence for Influenza Research and Surveillance (CEIRS) [HHSN272201400008C]
- Cohen Foundation
- JPB Foundation
- Open Philanthropy Project [2020-215611 [5384]]
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This study evaluated the correlation of six serological assays with authentic virus neutralization for SARS-CoV-2. The results showed that luciferase- and SEAP-expressing pseudotyped virus neutralizations were the most accurate in predicting authentic virus neutralization, followed by GFP-expressing pseudotyped virus neutralization, and then ELISAs. This research outlines the advantages and disadvantages of different assays, providing insights into the optimal surrogate assay for authentic virus neutralization and enhancing the accuracy of assessing protective immunity against SARS-CoV-2 post infection and vaccination.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in late 2019 and has since caused a global pandemic resulting in millions of cases and deaths. Diagnostic tools and serological assays are critical for controlling the outbreak, especially assays designed to quantitate neutralizing antibody levels, considered the best correlate of protection. As vaccines become increasingly available, it is important to identify reliable methods for measuring neutralizing antibody responses that correlate with authentic virus neutralization but can be performed outside biosafety level 3 (BSL3) laboratories. While many neutralizing assays using pseudotyped virus have been developed, there have been few studies comparing the different assays to each other as surrogates for authentic virus neutralization. Here, we characterized three enzyme-linked immunosorbent assays (ELISAs) and three pseudotyped vesicular stomatitis virus (VSV) neutralization assays and assessed their concordance with authentic virus neutralization. The most accurate assays for predicting authentic virus neutralization were luciferase- and secreted embryonic alkaline phosphatase (SEAP)-expressing pseudotyped virus neutralizations, followed by green fluorescent protein (GFP)-expressing pseudotyped virus neutralization, and then the ELISAs. IMPORTANCE The ongoing COVID-19 pandemic is caused by infection with severe acute respiratory syndrome virus 2 (SARS-CoV-2). Prior infection or vaccination can be detected by the presence of antibodies in the blood. Antibodies in the blood are also considered to be protective against future infections from the same virus. The gold standard assay for detecting protective antibodies against SARS-CoV-2 is neutralization of authentic SARSCoV-2 virus. However, this assay can only be performed under highly restrictive biocontainment conditions. We therefore characterized six antibody-detecting assays for their correlation with authentic virus neutralization. The significance of our research is in outlining the advantages and disadvantages of the different assays and identifying the optimal surrogate assay for authentic virus neutralization. This will allow for more accurate assessments of protective immunity against SARS-CoV-2 following infection and vaccination.
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