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A rapid real-time polymerase chain reaction-based live virus microneutralization assay for detection of neutralizing antibodies against SARS-CoV-2 in blood/serum

2021Artigo científico Acesso aberto
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dc.contributor.authorAbidi, Syed Hani
dc.contributor.authorImtiaz, Kehkashan
dc.contributor.authorKanji, Akbar
dc.contributor.authorQaiser, Shama
dc.contributor.authorKhan, Erum
dc.contributor.authorIqbal, Kiran
dc.contributor.authorVeldhoen, Marc
dc.contributor.authorGhias, Kulsoom
dc.contributor.authorSimas, J Pedro
dc.contributor.authorHasan, Zahra
dc.date.accessioned2022-01-05T11:27:27Z
dc.date.available2022-01-05T11:27:27Z
dc.date.issued2021
dc.descriptionCopyright: © 2021 Abidi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.pt_PT
dc.description.abstractBackground: Individuals recovering from COVID-19 are known to have antibodies against the Spike and other structural proteins. Antibodies against Spike have been shown to display viral neutralization. However, not all antibodies against Spike have neutralizing ability although they may be cross-reactive. There is a need for easy-to-use SARS-CoV-2 neutralizing assays for the determination of virus-neutralizing activity in sera of individuals. Here we describe a PCR-based micro-neutralization assay that can be used to evaluate the viral neutralization titers of serum from SARS-CoV-2 infected individuals. Methods: The SARS-CoV-2 strain used was isolated from a nasopharyngeal specimen of a COVID-19 case. The limiting dilution method was used to obtain a 50% tissue culture infective dose (TCID50) of Vero cells. For the micro-neutralization assay, 19 serum samples, with positive IgG titers against Spike Receptor-Binding Domain (RBD) were tested. After 24 hours, infected cells were inspected for the presence of a cytopathic effect, lysed and RNA RT-PCR conducted for SARS-CoV-2. PCR target Ct values were used to calculate percent neutralization/inhibition of SARS-CoV-2. Results: Out of 19 samples, 13 samples gave 100% neutralization at all dilutions, 1 sample showed neutralization at the first dilution, 4 samples showed neutralization at lower dilutions, while one sample did not demonstrate any neutralization. The RBD ODs and neutralization potential percentages were found to be positively correlated. Conclusion: We describe a rapid RT-PCR-based SARS-CoV-2 microneutralization assay for the detection of neutralizing antibodies. This can effectively be used to test the antiviral activity of serum antibodies for the investigation of both disease-driven and vaccine-induced responses.pt_PT
dc.description.sponsorshipThis study received funding support through a grant from the Provosts Academic Priorities Fund, The Aga Khan University. MV was supported by the European Union H2020 ERA project (No 667824 – EXCELLtoINNOV).pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationPLoS One. 2021 Dec 10;16(12):e0259551pt_PT
dc.identifier.doi10.1371/journal.pone.0259551pt_PT
dc.identifier.eissn1932-6203
dc.identifier.urihttp://hdl.handle.net/10451/50701
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherPLOSpt_PT
dc.relationLeverage research EXCELLence and INNOVation potential of Instituto de Medicina Molecular (IMM) through translational biomedical research in immunity and infection.
dc.relation.publisherversionhttps://journals.plos.org/plosone/pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.titleA rapid real-time polymerase chain reaction-based live virus microneutralization assay for detection of neutralizing antibodies against SARS-CoV-2 in blood/serumpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleLeverage research EXCELLence and INNOVation potential of Instituto de Medicina Molecular (IMM) through translational biomedical research in immunity and infection.
oaire.awardURIinfo:eu-repo/grantAgreement/EC/H2020/667824/EU
oaire.citation.issue12pt_PT
oaire.citation.titlePLOS ONEpt_PT
oaire.citation.volume16pt_PT
oaire.fundingStreamH2020
person.familyNameVeldhoen
person.familyNameSimas
person.givenNameMarc
person.givenNameJ Pedro
person.identifierhttps://scholar.google.pt/citations?hl=en&user=7vG1jLIAAAAJ
person.identifier.ciencia-id6413-A3A0-C7AD
person.identifier.ciencia-id3513-BF02-52B7
person.identifier.orcid0000-0002-1478-9562
person.identifier.orcid0000-0001-6982-9253
person.identifier.scopus-author-id8323789000
person.identifier.scopus-author-id7003457329
project.funder.identifierhttp://doi.org/10.13039/501100008530
project.funder.nameEuropean Commission
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublicationb6ddef66-b6bb-4ce5-b68a-e4b211aa2411
relation.isAuthorOfPublication28052826-b4b3-4e0d-9918-e0d115e3ff0b
relation.isAuthorOfPublication.latestForDiscovery28052826-b4b3-4e0d-9918-e0d115e3ff0b
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