Exploring low serum volume assays for SARS-CoV-2 antibodies

In a recent study posted to the medRxiv* preprint server, researchers developed several non-commercial (in-house) low serum volume assays for detecting and characterizing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies.

Study: Development and evaluation of low-volume tests to detect and characterise antibodies to SARS-CoV-2. Image Credit: Kateryna Kon/Shutterstock
Study: Development and evaluation of low-volume tests to detect and characterise antibodies to SARS-CoV-2. Image Credit: Kateryna Kon/Shutterstock

This news article was a review of a preliminary scientific report that had not undergone peer-review at the time of publication. Since its initial publication, the scientific report has now been peer reviewed and accepted for publication in a Scientific Journal. Links to the preliminary and peer-reviewed reports are available in the Sources section at the bottom of this article. View Sources

Antibody testing is important to assess SARS-CoV-2 seroprevalence, especially to identify asymptomatic cases among individuals residing in remote locations and low- and middle-income nations. Current coronavirus disease 2019 (COVID-19) diagnostic assays are expensive and technique-sensitive, warranting the need for cheap and simple tests for anti-SARS-CoV-2 antibodies.

About the study

In the present study, researchers developed low-serum volume (<10 µl) assays for COVID-19 diagnosis and optimizing discrimination between pre-pandemic controls and COVID-19 cases.

Serum samples were obtained from 984 pre-pandemic (known negative) individuals and from 269 suspected or polymerase chain reaction (PCR)-confirmed COVID-19 (known positive) patients.

The assays included total antibody and isotype-specific, standardized, low serum volume assays such as enzyme-linked immunosorbent assays (ELISA) for detecting total or antibody isotypes to SARS-CoV-2 nucleocapsid (N) protein or spike (S) protein and its receptor-binding domain (RBD). They also developed anti-RBD isotype-specific luciferase immunoprecipitation system (LIPS) assays and a novel S-RBD bridging LIPS total-antibody assay for COVID-19 diagnosis.

For every antigen, ELISAs were performed to assess the total antibody or Pan Immunoglobulin G (IgG) titers using commercial anti-human IgG secondary antiserum assays that detect all the isotypes (Pan antibodies), or IgA, IgM, and IgG isotypes using class-specific antibodies. In addition, LIPS assays measuring RBD-specific antibodies were developed. To measure IgA and IgG isotypes, an unlabelled RBD competing with Nluc-labelled RBD was used. For the evaluation of total antibody titers with high affinity for RBD, the novel S-RBD bridging assay was used.

The assay thresholds were determined based on three criteria: 1) 99th percentile of pre-pandemic controls; 2) 98th percentile of pre-pandemic controls, and 3) highest Youden Index for achieving balanced specificity and sensitivity. The assay results were compared with that of neutralization assays based on the neutralizing titer of antibody (ND50) values.

Results

In the study, the team developed 12 ELISAs that detected total antibodies or antibody isotypes to SARS-CoV-2 N, or S RBD. In the blind analysis, the S-RBD bridging LIPS and the S pan ELISA demonstrated >92% and >97% sensitivity and specificity, respectively among SARs-CoV-2-positive samples after 21 days of symptom onset or polymerase chain reaction (PCR) results.  

For every antigen, the Pan ELISAs performed better than the IgG-specific assays to distinguish COVID-19 cases from the pre-pandemic controls. SARS-CoV-2-positive cases demonstrated a greater area under the curve (AUCs) after the receiver operator characteristic (ROC) analyses.

In the IgA and IgG isotype titers, improved discrimination between the pre-pandemic samples and the COVID-19 samples was observed using Nluc-labelled RBD and unlabelled RBD whereas those observed in IgM titers were not optimal. As observed in the Pan ELISAs, the S-RBD bridging assay showed better AUCs post-ROC analysis for discriminating COVID-19 samples from the controls. Despite the same antigen use, the RBD LIPS assays demonstrated better AUCs than the RBD-specific ELISAs for all Ig isotypes.

In the threshold analysis, all assays accurately distinguished the COVID-19 cases from the pre-pandemic samples (AUCs between 0.95 to 0.997). The S-RBD bridging LIPS assay demonstrated optimal and near-perfect performance (AUC = 0.997). All assay thresholds showed >96% specificity. The S Pan ELISA demonstrated the highest specificity of 97.3%. In addition, the screening assays showed fewer intra-assay and inter-assay variations.

In the sensitivity analysis, the S Pan ELISA and the S-RBD bridging LIPS assays showed the highest sensitivities for COVID-19 detection (between 92.4 and 95.7% for >21 days post-infection). Compared to the commercial Roche assay, the S-RBD Bridging LIPS assay and the S Pan ELISA showed optimal sensitivity for COVID-19 detection, >93% sensitivity vs 85.5% for the Roche assay. The N Pan and RBD Pan ELISAs showed 76.97 and 76.3% sensitivity, respectively. The strongest agreements were observed between the S and RBD Pan ELISA assays and between the IgG and total antibody (Bridging or Pan) assays. The binding antibody units (BAU)/ml values in the in-house assays were comparable to those of the World Health Organization (WHO)/ National Institute for Biological Standards and Control (NIBSC) reference.

In the neutralization analyses, the assays reasonably neutralized the COVID-19 samples (ND50 >125) with results in the high positive range for the S and RBD-specific assays (e.g., > 0.72 Normalised OD on the S Pan assay and >20 units on the S-RBD Bridging assay). The S IgA and IgG ELISA assays showed the strongest correlation with half-maximal neutralization titers.

Overall, the study findings showed that low-volume assays are an economically viable option with high diagnostic accuracy for the detection and characterization of SARS-CoV-2 antibodies. Such assays would be highly beneficial for low and middle-income nations where commercial assays requiring 100-times higher serum volumes may not always be feasible.

This news article was a review of a preliminary scientific report that had not undergone peer-review at the time of publication. Since its initial publication, the scientific report has now been peer reviewed and accepted for publication in a Scientific Journal. Links to the preliminary and peer-reviewed reports are available in the Sources section at the bottom of this article. View Sources

Journal references:

Article Revisions

  • May 13 2023 - The preprint preliminary research paper that this article was based upon was accepted for publication in a peer-reviewed Scientific Journal. This article was edited accordingly to include a link to the final peer-reviewed paper, now shown in the sources section.
Pooja Toshniwal Paharia

Written by

Pooja Toshniwal Paharia

Pooja Toshniwal Paharia is an oral and maxillofacial physician and radiologist based in Pune, India. Her academic background is in Oral Medicine and Radiology. She has extensive experience in research and evidence-based clinical-radiological diagnosis and management of oral lesions and conditions and associated maxillofacial disorders.

Citations

Please use one of the following formats to cite this article in your essay, paper or report:

  • APA

    Toshniwal Paharia, Pooja Toshniwal Paharia. (2023, May 13). Exploring low serum volume assays for SARS-CoV-2 antibodies. News-Medical. Retrieved on November 07, 2024 from https://www.news-medical.net/news/20220509/Exploring-low-serum-volume-assays-for-SARS-CoV-2-antibodies.aspx.

  • MLA

    Toshniwal Paharia, Pooja Toshniwal Paharia. "Exploring low serum volume assays for SARS-CoV-2 antibodies". News-Medical. 07 November 2024. <https://www.news-medical.net/news/20220509/Exploring-low-serum-volume-assays-for-SARS-CoV-2-antibodies.aspx>.

  • Chicago

    Toshniwal Paharia, Pooja Toshniwal Paharia. "Exploring low serum volume assays for SARS-CoV-2 antibodies". News-Medical. https://www.news-medical.net/news/20220509/Exploring-low-serum-volume-assays-for-SARS-CoV-2-antibodies.aspx. (accessed November 07, 2024).

  • Harvard

    Toshniwal Paharia, Pooja Toshniwal Paharia. 2023. Exploring low serum volume assays for SARS-CoV-2 antibodies. News-Medical, viewed 07 November 2024, https://www.news-medical.net/news/20220509/Exploring-low-serum-volume-assays-for-SARS-CoV-2-antibodies.aspx.

Comments

The opinions expressed here are the views of the writer and do not necessarily reflect the views and opinions of News Medical.
Post a new comment
Post

While we only use edited and approved content for Azthena answers, it may on occasions provide incorrect responses. Please confirm any data provided with the related suppliers or authors. We do not provide medical advice, if you search for medical information you must always consult a medical professional before acting on any information provided.

Your questions, but not your email details will be shared with OpenAI and retained for 30 days in accordance with their privacy principles.

Please do not ask questions that use sensitive or confidential information.

Read the full Terms & Conditions.

You might also like...
Study finds nirmatrelvir-ritonavir reduces severe COVID-19 and long COVID risks in high-risk patients