☆ 4.5 Article

Development and characterization of a quantitative ELISA to detect anti-SARS-CoV-2 spike antibodies

HELIYON (2021)

Journal

HELIYON

Volume 7, Issue 12, Pages -

Publisher

ELSEVIER SCI LTD

DOI: 10.1016/j.heliyon.2021.e08444

Keywords

Coronavirus disease 2019; Severe acute respiratory syndrome coronavirus-; 2 (SARSCoV-2); Antibody tests; Diagnosis of COVID-19; ELISA; IgG antibody assay

Funding

JPB Foundation
Open Philanthropy Project [2020-215611]

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Abstract

The novel clinical assay developed for the detection and quantitation of antibodies against SARS-CoV-2 showed high sensitivity and specificity, with sensitivity of 92.5% and specificity of 100%. This assay could be widely used in clinical settings to evaluate SARS-CoV-2 seroprevalence and antibody profiling in different populations.

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 coronavirus disease 2019 (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.

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Performance assessment of seven SARS-CoV-2 IgG enzyme-linked immunosorbent assays

Ketki Deshpande, Ullas Pt, Ojas Kaduskar, Neetu Vijay, Aparna Rakhe, Shankar Vidhate, Kirtee Khutwad, Gururaj Rao Deshpande, Bipin Tilekar, Sanskruti Saka, Kshitija Gadekar, Roshni Patil, Pragya Yadav, Varsha Potdar, Yogesh Gurav, Priyanka Gupta, Harmanmeet Kaur, Jitendra Narayan, Gajanan Sapkal, Priya Abraham

Summary: Seven commercially available ELISA kits demonstrated high accuracy in detecting IgG antibodies against SARS-CoV-2, with DiaPro COVID-19 IgG ELISA showing the highest sensitivity. These assays can be valuable in tracing mild infections, cohort studies, and pre-vaccine evaluations due to their good accuracy indices.

JOURNAL OF MEDICAL VIROLOGY (2021)