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Development of a high throughput SARS-CoV-2 antibody testing pathway using dried blood spot specimens

Moat, Stuart James, Zelek, Wiola M., Carne, Emily, Ponsford, Mark J., Bramhall, Kathyryn, Jones, Sara, El-Shanawany, Tariq, Wise, Matthew J., Thomas, Annette, George, Chloe, Fegan, Christopher ORCID:, Steven, Rachael, Webb, Russell, Weeks, Ian ORCID:, Morgan, Brian Paul and Jolles, Stephen 2021. Development of a high throughput SARS-CoV-2 antibody testing pathway using dried blood spot specimens. Annals of Clinical Biochemistry 58 (2) 10.1177/0004563220981106

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Background: Serologic assays for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) have roles in seroepidemiology, convalescent plasma-testing, antibody durability and vaccine studies. Currently, SARS-CoV-2 serology is performed using serum/plasma collected by venepuncture. Dried bloodspot (DBS) testing offers significant advantages; as it is minimally invasive, avoids venepuncture with specimens being mailed to the laboratory. Methods: A pathway utilising a newborn screening laboratory infrastructure was developed using an Enzyme-Linked Immunosorbent assay (ELISA) to detect IgG antibodies against the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein in DBS specimens. Paired plasma and DBS specimens from SARS-CoV-2 antibody positive and negative subjects and PCR positive subjects were tested. DBS specimen stability, effect of blood volume and punch location were also evaluated. Results: DBS from antibody-negative (n=85) and positive (n=35) subjects and PCR positive subjects (n=11) had a mean (SD; range) optical density (OD) of 0.14 (0.046; 0.03-0.27), 0.98 (0.41; 0.31-1.64) and 1.12 (0.37; 0.49-1.54), respectively. An action value OD >0.28 correctly assigned all cases. The weighted Deming regression for comparison of the DBS and the plasma assay yielded: y=0.004041+1.005x, r=0.991, Sy/x 0.171, n=82. Extraction efficiency of antibodies from DBS was >99%. DBS were stable for at least 28 days at ambient room temperature and humidity. Conclusions: SARS-CoV-2 IgG RBD antibodies can be reliably detected in DBS. DBS serological testing offers lower costs than either point of care or serum/plasma assays that require patient travel, phlebotomy and hospital/clinic resources; the development of a DBS assay may be particularly important for resource poor settings.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Additional Information: This article is distributed under the terms of the Creative Commons Attribution 4.0 License (
Publisher: SAGE Publications (UK and US)
ISSN: 0004-5632
Date of First Compliant Deposit: 7 December 2020
Date of Acceptance: 24 September 2020
Last Modified: 26 Nov 2023 16:18

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