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Detection of SARS-CoV-2 antibodies in serum and dried blood spot samples of vaccinated individuals using a sensitive homogeneous proximity extension assay
Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Sweden; Unit of Affinity Proteomics Uppsala, Science for Life Laboratory, Sweden.
Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Sweden.
Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Sweden.
Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Sweden; Unit of Affinity Proteomics Uppsala, Science for Life Laboratory, Sweden.
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2022 (English)In: New Biotechnology, ISSN 1871-6784, E-ISSN 1876-4347, Vol. 72, p. 139-148Article in journal (Refereed) Published
Abstract [en]

A homogeneous PCR-based assay for sensitive and specific detection of antibodies in serum or dried blood spots (DBS) is presented and the method is used to monitor individuals infected with or vaccinated against SARS-CoV-2. Detection probes were prepared by conjugating the recombinant spike protein subunit 1 (S1), containing the receptor binding domain (RBD) of SARS-CoV-2, to each of a pair of specific oligonucleotides. The same was done for the nucleocapsid protein (NP). Upon incubation with serum or DBS samples, the bi- or multivalency of the antibodies (IgG, IgA or IgM) brings pairs of viral proteins with their conjugated oligonucleotides in proximity, allowing the antibodies to be detected by a modified proximity extension assay (PEA). Anti-S1 and anti-NP antibodies could be detected simultaneously from one incubation reaction. This Antibody PEA (AbPEA) test uses only 1 µl of neat or up to 100,000-fold diluted serum or one ø1.2 mm disc cut from a DBS. All 100 investigated sera and 21 DBS collected prior to the COVID-19 outbreak were negative, demonstrating a 100% specificity. The area under the curve, as evaluated by Receiver Operating Characteristic (ROC) analysis reached 0.998 (95%CI: 0.993–1) for samples taken from 11 days after symptoms onset. The kinetics of antibody responses were monitored after a first and second vaccination using serially collected DBS from 14 individuals. AbPEA offers highly specific and sensitive solution-phase antibody detection without requirement for secondary antibodies, no elution step when using DBS sample in a simple procedure that lends itself to multiplex survey of antibody responses.

Place, publisher, year, edition, pages
Elsevier, 2022. Vol. 72, p. 139-148
Keywords [en]
Finger-prick dried blood spot, Homogenous serological assay, Multiplex immunoassay, PCR-based antibody detection, Proximity extension assay, SARS-CoV-2 antibody
National Category
Infectious Medicine
Identifiers
URN: urn:nbn:se:umu:diva-201459DOI: 10.1016/j.nbt.2022.11.004ISI: 000896515300004PubMedID: 36423830Scopus ID: 2-s2.0-85142674902OAI: oai:DiVA.org:umu-201459DiVA, id: diva2:1716698
Funder
Knut and Alice Wallenberg Foundation, 2020.0182Swedish Foundation for Strategic Research, SB16-0046Swedish Research Council, 2020-02258Available from: 2022-12-06 Created: 2022-12-06 Last updated: 2023-09-05Bibliographically approved

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Monsen, Tor J.Mei, Ya-Fang

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