Open AccessAn Assessment of Serological Assays for SARS-CoV-2 as Surrogates for Authentic Virus Neutralization
Author(s) -
Nicholas Wohlgemuth,
Kendall Whitt,
Sean Cherry,
Ericka Kirkpatrick Roubidoux,
Chun-Yang Lin,
Kim Allison,
Ashleigh Gowen,
Pamela Freiden,
E. Kaitlynn Allen,
St. Jude Investigative Team,
Aditya H. Gaur,
Jeremie H. Estepp,
Tang Li,
Tomi Mori,
Diego R. Hijano,
Hana Hakim,
Maureen A. McGargill,
Florian Krammer,
Michael A. Whitt,
Joshua Wolf,
Paul G. Thomas,
Stacey SchultzCherry
Publication year - 2021
Publication title -
microbiology spectrum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.502
H-Index - 51
ISSN - 2165-0497
DOI - 10.1128/spectrum.01059-21
Subject(s) - neutralization , virology , virus , vesicular stomatitis virus , serology , antibody , mononegavirales , biology , ebola virus , neutralizing antibody , viral disease , immunology , paramyxoviridae
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 SARS-CoV-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.