Dataset Information

Detection of neutralizing antibodies against multiple SARS-CoV-2 strains in dried blood spots using cell-free PCR.

ABSTRACT: An easily implementable serological assay to accurately detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) neutralizing antibodies is urgently needed to better track herd immunity, vaccine efficacy and vaccination rates. Herein, we report the Split-Oligonucleotide Neighboring Inhibition Assay (SONIA) which uses real-time qPCR to measure the ability of neutralizing antibodies to block binding between DNA-barcoded viral spike protein subunit 1 and the human angiotensin-converting enzyme 2 receptor protein. The SONIA neutralizing antibody assay using finger-prick dried blood spots displays 91-97% sensitivity and 100% specificity in comparison to the live-virus neutralization assays using matched serum specimens for multiple SARS-CoV-2 variants-of-concern. The multiplex version of this neutralizing antibody assay, using easily collectable finger-prick dried blood spots, can be a valuable tool to help reveal the impact of age, pre-existing health conditions, waning immunity, different vaccination schemes and the emergence of new variants-of-concern.

SUBMITTER: Danh K

PROVIDER: S-EPMC9302227 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Detection of neutralizing antibodies against multiple SARS-CoV-2 strains in dried blood spots using cell-free PCR.

Danh Kenneth K Karp Donna Grace DG Singhal Malvika M Tankasala Akshaya A Gebhart David D de Jesus Cortez Felipe F Tandel Devangkumar D Robinson Peter V PV Seftel David D Stone Mars M Simmons Graham G Bagri Anil A Schreiber Martin A MA Buser Andreas A Holbro Andreas A Battegay Manuel M Morris Mary Kate MK Hanson Carl C Mills John R JR Granger Dane D Theel Elitza S ES Stubbs James R JR Corash Laurence M LM Tsai Cheng-Ting CT

Nature communications 20220721 1

An easily implementable serological assay to accurately detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) neutralizing antibodies is urgently needed to better track herd immunity, vaccine efficacy and vaccination rates. Herein, we report the Split-Oligonucleotide Neighboring Inhibition Assay (SONIA) which uses real-time qPCR to measure the ability of neutralizing antibodies to block binding between DNA-barcoded viral spike protein subunit 1 and the human angiotensin-converting ...[more]

PMID: 35864104

Similar Datasets

Project description:In the earliest days of the COVID-19 pandemic, the collection of dried blood spots (DBS) enabled public health laboratories to undertake population-scale seroprevalence studies used to estimate rates of SARS-CoV-2 exposure. With SARS-CoV-2 seropositivity levels now estimated to exceed 94% in the United States, attention has turned to using DBS to assess neutralizing antibodies within cohorts of interest. With this goal in mind, we generated contrived DBS (cDBS) and whole blood-derived DBS from convalescent and vaccinated individuals and subjected DBS eluates to a battery of assays, including a SARS-CoV-2 multiplexed microsphere immunoassay (MIA), a receptor binding domain (RBD)-human ACE2 inhibition assay (iACE2), a cell-based pseudovirus neutralization assay, and real-time PCR-based surrogate neutralization assay (NAB-Sure). The DBS results were benchmarked against paired serum samples tested in a clinically validated SARS-CoV-2 plaque reduction neutralization titer (PRNT) assay. The results of an 8-plex MIA and NAB-Sure assays demonstrated highly significant correlations with PRNT values when evaluated with a panel of 86 paired serum-DBS samples. Both the MIA and NAB-Sure are adaptable to automated liquid handlers for high-throughput capacity. While neutralizing assays were limited to the ancestral SARS-CoV-2 WA1, this study nonetheless represents an important proof of concept demonstrating the potential utility of DBS as a biospecimen type for use in assessing immunity to SARS-CoV-2 at the community and population levels.IMPORTANCESARS-CoV-2 variants of concern continue to circulate globally and remain a serious health threat to large segments of the population. From a public health standpoint, identifying vulnerable communities based on immune status is critical in terms of vaccine booster recommendations. In this report, we investigated the utility of dried blood spots (DBS) as a biospecimen type from which to estimate SARS-CoV-2 neutralizing antibody titers. Using contrived and whole blood-derived DBS, we demonstrate that SARS-CoV-2 neutralizing antibodies are readily measurable in DBS eluates and correlate with plaque reduction neutralization titer (PRNT) values from paired serum samples. Moreover, several of the methods used to estimate SARS-CoV-2 neutralizing antibodies in DBS eluates are adaptable to high-throughput platforms.

Project description:Dried blood spots (DBS) are broadly used for different serological analyses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody assessment. In order to biobank DBS samples, an understanding of the preservation of SARS-CoV-2 antibodies is needed. Therefore, we assessed the stability of SARS-CoV-2 antibodies on DBS during short- and long-term storage under different storage temperatures. Seven sample donors were enrolled, each donating twenty 6 mm DBS to assess anti-spike (S1) SARS-CoV-2 IgG antibodies (EUROIMMUN). Baseline samples were analyzed on the day of collection. The remainder of the samples was stored in grip seal bags kept in a cryobox at room temperature/4°C until 2 months after collection and at -20°C until 2 years after collection. Samples were analyzed at regular intervals within the total storage duration and after one and five freeze-thaw cycles. A pooled coefficient of variation was calculated for each storage temperature. We found that anti-S1 SARS-CoV-2 antibodies collected on DBS saver cards remain stable during short-term storage at RT, 4°C, and -20°C (at least to 2 months) and long-term storage at -20°C (at least 2 years). Moreover, up to five freeze-thaw cycles can occur without impacting the anti-S1 SARS-CoV-2 antibody level. The inter-assay coefficient of variation lies between 10 and 15%. As DBS can be preserved for both shorter periods of time at RT and longer periods of time at -20°C, they are a perfect application for studies that require sample shipment by mail, self-sampling studies, studies in limited resource settings and biobanking.IMPORTANCEDried blood spots (DBS) are currently widely used as a microsampling technique for different qualitative and quantitative serological assessments. Yet, there is a lack of long-term stability and storage condition studies. In our study, first, we assessed the stability of SARS-CoV-2 antibodies on DBS up to 2 years post-collection. We believe that our data are not only important for future COVID-19 research but also for studies on other infections/diseases using DBS-based serology.

Dataset Information

Detection of neutralizing antibodies against multiple SARS-CoV-2 strains in dried blood spots using cell-free PCR.

Publications

Detection of neutralizing antibodies against multiple SARS-CoV-2 strains in dried blood spots using cell-free PCR.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets