Validation of a SARS-CoV-2 spike protein ELISA for use in contact investigations and sero-surveillance

  1. Brandi Freeman
  2. Sandra Lester
  3. Lisa Mills
  4. Mohammad Ata Ur Rasheed
  5. Stefany Moye
  6. Olubukola Abiona
  7. Geoffrey B. Hutchinson
  8. Maria Morales-Betoulle
  9. Inna Krapinunaya
  10. Ardith Gibbons
  11. Cheng-Feng Chiang
  12. Deborah Cannon
  13. John Klena
  14. Jeffrey A. Johnson
  15. Sherry Michele Owen
  16. Barney S. Graham
  17. Kizzmekia S. Corbett
  18. Natalie J. Thornburg

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Abstract

Since emergence of SARS-CoV-2 in late 2019, there has been a critical need to understand prevalence, transmission patterns, to calculate the burden of disease and case fatality rates. Molecular diagnostics, the gold standard for identifying viremic cases, are not ideal for determining true case counts and rates of asymptomatic infection. Serological detection of SARS-CoV-2 specific antibodies can contribute to filling these knowledge gaps. In this study, we describe optimization and validation of a SARS-CoV-2-specific-enzyme linked immunosorbent assay (ELISA) using the prefusion-stabilized form of the spike protein [1]. We performed receiver operator characteristic (ROC) analyses to define the specificities and sensitivities of the optimized assay and examined cross reactivity with immune sera from persons confirmed to have had infections with other coronaviruses. These assays will be used to perform contact investigations and to conduct large-scale, cross sectional surveillance to define disease burden in the population.

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  1. ScreenIT

    SciScore for 10.1101/2020.04.24.057323: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    HRP-conjugated goat anti-human antibodies (KPL) diluted at 1:2000 in serum diluent were added to washed plates and incubated at 37°C in a humidified chamber for 1 hour.
    anti-human antibodies (KPL
    suggested: (SeraCare KPL Cat# 5210-0158, RRID:AB_2773725)
    Experimental Models: Cell Lines
    SentencesResources
    ELISA: The pre-fusion stabilized ectodomain of SARS-CoV-2 spike (S) was expressed in suspension adapted HEK-293 cells as previously described [1].
    HEK-293
    suggested: CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045)
    Software and Algorithms
    SentencesResources
    Statistics were performed using GraphPad Prism software (7.04).
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.1101/2020.04.24.057323v2 on bioRxiv
  3. Version published to 10.1101/2020.04.24.057323v1 on bioRxiv