Standardization of ELISA protocols for serosurveys of the SARS-CoV-2 pandemic using clinical and at-home blood sampling

  1. Carleen Klumpp-Thomas
  2. Heather Kalish
  3. Matthew Drew
  4. Sally Hunsberger
  5. Kelly Snead
  6. Michael P. Fay
  7. Jennifer Mehalko
  8. Anandakumar Shunmugavel
  9. Vanessa Wall
  10. Peter Frank
  11. John-Paul Denson
  12. Min Hong
  13. Gulcin Gulten
  14. Simon Messing
  15. Jennifer Hicks
  16. Sam Michael
  17. William Gillette
  18. Matthew D. Hall
  19. Matthew J. Memoli
  20. Dominic Esposito
  21. Kaitlyn Sadtler

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Abstract

The extent of SARS-CoV-2 infection throughout the United States population is currently unknown. High quality serology is key to avoiding medically costly diagnostic errors, as well as to assuring properly informed public health decisions. Here, we present an optimized ELISA-based serology protocol, from antigen production to data analyses, that helps define thresholds for IgG and IgM seropositivity with high specificities. Validation of this protocol is performed using traditionally collected serum as well as dried blood on mail-in blood sampling kits. Archival (pre-2019) samples are used as negative controls, and convalescent, PCR-diagnosed COVID-19 patient samples serve as positive controls. Using this protocol, minimal cross-reactivity is observed for the spike proteins of MERS, SARS1, OC43 and HKU1 viruses, and no cross reactivity is observed with anti-influenza A H1N1 HAI. Our protocol may thus help provide standardized, population-based data on the extent of SARS-CoV-2 seropositivity, immunity and infection.

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  1. Version published to 10.1038/s41467-020-20383-x
  2. ScreenIT

    SciScore for 10.1101/2020.05.21.20109280: (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
    Secondary antibody (HRP conjugated: Goat anti-Human IgG (H+L) Cross-Adsorbed Secondary Antibody,
    anti-Human IgG
    suggested: None
    , Goat anti-Human IgM Cross-Adsorbed Secondary Antibody, Goat anti-Human IgA Cross-Adsorbed Secondary Antibody; ThermoFisher) was diluted at 1:4000 in blocking buffer and 100 μl of each antibody was then added to each well and incubated for 1 hour.
    anti-Human IgM
    suggested: None
    anti-Human IgA
    suggested: None
    These samples were tested for the presence of SARS-CoV-2 antibody and were also run against a panel of spike proteins from four other beta-coronaviruses; MERS, SARS1, OC43 and HKU1 to establish seroprevalence estimates for these pre-pandemic coronaviruses in a current population.
    SARS1
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Protein expression: Manufacturer’s protocols were followed for the transfection and culturing of Expi293 cells (Thermo Fisher Scientific, Waltham MA)
    Expi293
    suggested: RRID:CVCL_D615)

    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: We found the following clinical trial numbers in your paper:

    IdentifierStatusTitle
    NCT04334954Active, not recruitingSARS-COV2 Pandemic Serosurvey and Blood Sampling
    NCT01386424RecruitingScreening for LID Clinical Studies Unit Healthy Volunteer Pr…

    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.

  3. Version published to 10.1101/2020.05.21.20109280 on medRxiv