tetano
Editor, Senior Moderator
Heliyon
. 2021 Nov 20;e08444.
doi: 10.1016/j.heliyon.2021.e08444. Online ahead of print.
Development and characterization of a quantitative ELISA to detect anti-SARS-CoV-2 spike antibodies
Magdalena M Żak[SUP] 1 [/SUP], Aryeh Stock[SUP] 2 [/SUP], Daniel Stadlbauer[SUP] 3 4 [/SUP], Wei Zhang[SUP] 2 [/SUP], Kirstie Cummings[SUP] 5 [/SUP], William Marsiglia[SUP] 6 [/SUP], Arsen Zargarov[SUP] 2 [/SUP], Fatima Amanat[SUP] 3 [/SUP], Monica Tamayo[SUP] 2 [/SUP], Carlos Cordon-Cardo[SUP] 2 [/SUP], Florian Krammer[SUP] 3 [/SUP], Damodara Rao Mendu[SUP] 2 [/SUP]
Affiliations
Abstract
A novel clinical assay for the detection and quantitation of antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was adapted from an in-house, research-based enzyme-linked immunosorbent assay (ELISA). Development and validation were performed under regulatory guidelines, and the test obtained emergency use authorization (EUA) from the New York State Department of Health (NYSDOH) and the Food and Drug Administration (FDA). The Mount Sinai COVID-19 antibody assay is an orthogonal, quantitative direct ELISA test which detects antibodies reactive to the receptor binding domain (RBD) and the spike protein of the novel SARS-CoV-2. The assay is performed on 96-well plates coated with either SARS-CoV-2 recombinant RBD or spike proteins. The test is divided into two stages, a qualitative screening assay against RBD and a quantitative assay against the full-length spike protein. The test uses pooled high titer serum as a reference standard. Negative pre-COVID-19 and positive post-COVID-19, PCR-confirmed specimens were incorporated in each ELISA test run, and the assays were performed independently at two different locations. The Mount Sinai COVID-19 serology performed with high sensitivity and specificity, 92.5% (95% CI: 0.785 - 0.980) and 100% (CI: 0.939 - 1.000) respectively. Between-run precision was assessed with a single run repeated over 22 days; and within-run precision was assessed with 10 replicates per day over 22 days. Both were within reported acceptance criteria (CV ≤20%). This population-based study reveals the applicability and reliability of this novel orthogonal COVID-19 serology test for the detection and quantitation of antibodies against SARS-CoV-2, allowing a broad set of clinical applications, including the broad evaluation of SARS-CoV-2 seroprevalence and antibody profiling in different population subsets.
Keywords: Antibody tests; Coronavirus disease 2019; Diagnosis of COVID-19; ELISA; IgG antibody assay; Severe acute respiratory syndrome coronavirus-2 (SARSCoV-2).
. 2021 Nov 20;e08444.
doi: 10.1016/j.heliyon.2021.e08444. Online ahead of print.
Development and characterization of a quantitative ELISA to detect anti-SARS-CoV-2 spike antibodies
Magdalena M Żak[SUP] 1 [/SUP], Aryeh Stock[SUP] 2 [/SUP], Daniel Stadlbauer[SUP] 3 4 [/SUP], Wei Zhang[SUP] 2 [/SUP], Kirstie Cummings[SUP] 5 [/SUP], William Marsiglia[SUP] 6 [/SUP], Arsen Zargarov[SUP] 2 [/SUP], Fatima Amanat[SUP] 3 [/SUP], Monica Tamayo[SUP] 2 [/SUP], Carlos Cordon-Cardo[SUP] 2 [/SUP], Florian Krammer[SUP] 3 [/SUP], Damodara Rao Mendu[SUP] 2 [/SUP]
Affiliations
- PMID: 34841098
- PMCID: PMC8605824
- DOI: 10.1016/j.heliyon.2021.e08444
Abstract
A novel clinical assay for the detection and quantitation of antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was adapted from an in-house, research-based enzyme-linked immunosorbent assay (ELISA). Development and validation were performed under regulatory guidelines, and the test obtained emergency use authorization (EUA) from the New York State Department of Health (NYSDOH) and the Food and Drug Administration (FDA). The Mount Sinai COVID-19 antibody assay is an orthogonal, quantitative direct ELISA test which detects antibodies reactive to the receptor binding domain (RBD) and the spike protein of the novel SARS-CoV-2. The assay is performed on 96-well plates coated with either SARS-CoV-2 recombinant RBD or spike proteins. The test is divided into two stages, a qualitative screening assay against RBD and a quantitative assay against the full-length spike protein. The test uses pooled high titer serum as a reference standard. Negative pre-COVID-19 and positive post-COVID-19, PCR-confirmed specimens were incorporated in each ELISA test run, and the assays were performed independently at two different locations. The Mount Sinai COVID-19 serology performed with high sensitivity and specificity, 92.5% (95% CI: 0.785 - 0.980) and 100% (CI: 0.939 - 1.000) respectively. Between-run precision was assessed with a single run repeated over 22 days; and within-run precision was assessed with 10 replicates per day over 22 days. Both were within reported acceptance criteria (CV ≤20%). This population-based study reveals the applicability and reliability of this novel orthogonal COVID-19 serology test for the detection and quantitation of antibodies against SARS-CoV-2, allowing a broad set of clinical applications, including the broad evaluation of SARS-CoV-2 seroprevalence and antibody profiling in different population subsets.
Keywords: Antibody tests; Coronavirus disease 2019; Diagnosis of COVID-19; ELISA; IgG antibody assay; Severe acute respiratory syndrome coronavirus-2 (SARSCoV-2).