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Detection of SARS-CoV-2 IgG antibodies in dried blood spots.

ABSTRACT: The COVID-19 pandemic led to development of numerous serologic tests. To obviate the need for phlebotomy services, we validated dried blood spots (DBS) for anti-SARS-CoV-2 serologic testing. Immunoglobulins were extracted from 3 mm DBS punches and the extracts were analyzed using the Euroimmun anti-SARS-CoV-2 IgG ELISA. Various pre-analytical factors were studied. Results were favorable for DBS stored for at least 67 days at or below 37°C. Comparison between paired serum and DBS specimens tested by the Euroimmun ELISA showed 96.8% and 81.3% positive and negative agreement, respectively, indicating that confirmatory testing of positive Euroimmun results on DBS extracts is necessary to achieve clinical accuracy. Our findings suggest that any SARS-CoV-2 antibody assay that requires pre-dilution of serum is amenable to DBS as an alternate specimen type that is relatively low cost, self-collectable, stable, can be shipped by standard mail and could be used to establish the seroprevalence of large populations.

SUBMITTER: Turgeon CT

PROVIDER: S-EPMC8116316 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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Detection of SARS-CoV-2 IgG antibodies in dried blood spots.

Turgeon Coleman T CT Sanders Karen A KA Granger Dane D Nett Stephanie L SL Hilgart Heather H Matern Dietrich D Theel Elitza S ES

Diagnostic microbiology and infectious disease 20210513 1

The COVID-19 pandemic led to development of numerous serologic tests. To obviate the need for phlebotomy services, we validated dried blood spots (DBS) for anti-SARS-CoV-2 serologic testing. Immunoglobulins were extracted from 3 mm DBS punches and the extracts were analyzed using the Euroimmun anti-SARS-CoV-2 IgG ELISA. Various pre-analytical factors were studied. Results were favorable for DBS stored for at least 67 days at or below 37°C. Comparison between paired serum and DBS specimens tested ...[more]

PMID: 34116343

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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 SARS-CoV-2 IgG antibodies in dried blood spots.

Publications

Detection of SARS-CoV-2 IgG antibodies in dried blood spots.

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