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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

2021-12-10Journal article Open access
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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.accessioned2021-12-22T15:33:35Z
dc.date.available2021-12-22T15:33:35Z
dc.date.issued2021-12-10
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.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1371/journal.pone.0259551pt_PT
dc.identifier.eid85121052729
dc.identifier.issn1932-6203
dc.identifier.pmcPMC8664206
dc.identifier.pmid34890401
dc.identifier.urihttp://hdl.handle.net/10400.14/36255
dc.identifier.wos000747293600003
dc.language.isoengpt_PT
dc.peerreviewedyespt_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.citation.titlePLoS ONEpt_PT
oaire.citation.volume16pt_PT
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT

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