An Assessment of Serological Assays for SARS-CoV-2 as Surrogates for Authentic Virus Neutralization

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Abstract

The ongoing COVID-19 pandemic is caused by infection with severe acute respiratory syndrome virus 2 (SARS-CoV-2). Prior infection or vaccination can be detected by the presence of antibodies in the blood.

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  1. SciScore for 10.1101/2021.04.14.21255399: (What is this?)

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

    Table 1: Rigor

    Ethicsnot detected.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    SARS-CoV-2 neutralizing antibody assay: Serially diluted plasma samples were mixed with diluted (approximately 6 PFU/cm2) SARS-CoV-2 (2019n-CoV/USA_WA1/2020) in EMEM supplemented with 5% FBS, 100 U/mL penicillin, 100 µg/mL streptomycin, and 2 mM GlutaMax.
    SARS-CoV-2
    suggested: None
    Virus was adsorbed for 1 hr, the inoculum was removed, cells were rinsed once with serum-free DMEM and then 4 ml of hybridoma supernatant containing the I1 monoclonal antibody24 was added for 30 minutes to neutralize residual VSV-ΔG pseudotyped virus from the inoculum and then replaced with DMEM containing 20% fetal bovine serum.
    antibody24
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Tissue culture: VeroE6 cells stably expressing TMPRSS2 (Vero-TMPRSS2) (XenoTech) were cultured in Eagle’s Minimal Essential Medium (EMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, 100 µg/mL streptomycin, and 2 mM GlutaMax (Gibco).
    VeroE6
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    After the incubation period, culture medium was removed from Vero-TMPRSS2 cells and virus/plasma mixture was added to the cells in triplicate.
    Vero-TMPRSS2
    suggested: JCRB Cat# JCRB1818, RRID:CVCL_YQ48)
    Baby hamster kidney (BHK-21) cells in 10 cm dishes were transfected using Lipofectamine 2000 according to the manufacturer’s instructions with 24 μg of a plasmid encoding a codon-optimized cDNA for the SARS-CoV-2 spike15, which was generously provided by Florian Krammer.
    BHK-21
    suggested: None
    Software and Algorithms
    SentencesResources
    AUC and ND50 values for the different assays were compared by mixed-effects model with the Geisser-Greenhouse correction and Tukey multiple comparisons post-test and p-value adjustment in GraphPad Prism (version 9.0.0).
    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.
    • Thank you for including a protocol registration statement.

    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.