Novel ELISA Protocol Links Pre-Existing SARS-CoV-2 Reactive Antibodies With Endemic Coronavirus Immunity and Age and Reveals Improved Serologic Identification of Acute COVID-19 via Multi-Parameter Detection

Frontiers in Immunology

Ian R. Rifkin¹  Amedeo J. Cappione²  Wenda Gao³  Gustavo Mostoslavsky⁴  Dylan Steiner⁵  Filiz Korkmaz⁵  Patricia Urick⁵  Lillia A. Baird⁵  Karen Quillen⁵  Jacob L. Berrigan⁶  Elizabeth Chavez⁶  Suryaram Gummuluru⁶  Rachel R. Yuen⁶  Jennifer E. Snyder-Cappione⁶  Erika L. Smith⁶  Yachana Kataria⁷  Anna C. Belkina⁷  Nahid Bhadelia⁹  Ronald B. Corley⁹  Riley M.F. Pihl^1,10  Nina H. Lin^1,12  Manish Sagar^1,12  Alex Olson^1,12 

[1] 0Renal Section, Department of Medicine, VA Boston Healthcare System, Boston, MA, United States;1MilliporeSigma, Burlington, MA, United States;2Antagen Pharmaceuticals, Boston, MA, United States;Center for Regenerative Medicine, Boston University School of Medicine, Boston, MA, United States;Department of Medicine, Boston University School of Medicine, Boston, MA, United States;Department of Microbiology, Boston University School of Medicine, Boston, MA, United States;Department of Pathology and Laboratory Medicine, Boston University School of Medicine, Boston, MA, United States;Flow Cytometry Core Facility, Boston University School of Medicine, Boston, MA, United States;National Emerging Infectious Diseases Laboratories (NEIDL), Boston University, Boston, MA, United States;PiBS Program, Boston University School of Medicine, Boston, MA, United States;Renal Section, Department of Medicine, Boston University School of Medicine, Boston, MA, United States;Section of Infectious Diseases, Department of Medicine, Boston Medical Center, Boston, MA, United States;

关键词: SARS-CoV-2;    COVID-19;    antibodies;    serology;    nucleocapsid (N);    receptor binding domain (RBD);   

DOI  :  10.3389/fimmu.2021.614676

来源: DOAJ

【 摘 要 】

The COVID-19 pandemic has drastically impacted work, economy, and way of life. Sensitive measurement of SARS-CoV-2 specific antibodies would provide new insight into pre-existing immunity, virus transmission dynamics, and the nuances of SARS-CoV-2 pathogenesis. To date, existing SARS-CoV-2 serology tests have limited utility due to insufficient reliable detection of antibody levels lower than what is typically present after several days of symptoms. To measure lower quantities of SARS-CoV-2 IgM, IgG, and IgA with higher resolution than existing assays, we developed a new ELISA protocol with a distinct plate washing procedure and timed plate development via use of a standard curve. Very low optical densities from samples added to buffer coated wells at as low as a 1:5 dilution are reported using this ‘BU ELISA’ method. Use of this method revealed circulating SARS-CoV-2 receptor binding domain (RBD) and nucleocapsid protein (N) reactive antibodies (IgG, IgM, and/or IgA) in 44 and 100 percent of pre-pandemic subjects, respectively, and the magnitude of these antibodies tracked with antibody levels of analogous viral proteins from endemic coronavirus (eCoV) strains. The disease status (HIV, SLE) of unexposed subjects was not linked with SARS-CoV-2 reactive antibody levels; however, quantities were significantly lower in subjects over 70 years of age compared with younger counterparts. Also, we measured SARS-CoV-2 RBD- and N- specific IgM, IgG, and IgA antibodies from 29 SARS-CoV-2 infected individuals at varying disease states, including 10 acute COVID-19 hospitalized subjects with negative serology results by the EUA approved Abbott IgG chemiluminescent microparticle immunoassay. Measurements of SARS-CoV-2 RBD- and N- specific IgM, IgG, IgA levels measured by the BU ELISA revealed higher signal from 9 of the 10 Abbott test negative COVID-19 subjects than all pre-pandemic samples for at least one antibody specificity/isotype, implicating improved serologic identification of SARS-CoV-2 infection via multi-parameter, high sensitive antibody detection. We propose that this improved ELISA protocol, which is straightforward to perform, low cost, and uses readily available commercial reagents, is a useful tool to elucidate new information about SARS-CoV-2 infection and immunity and has promising implications for improved detection of all analytes measurable by this platform.

【 授权许可】

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