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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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
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources 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-2suggested: NoneVirus 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 … 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
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources 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-2suggested: NoneVirus 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. antibody24suggested: NoneExperimental Models: Cell Lines Sentences Resources 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). VeroE6suggested: 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-TMPRSS2suggested: 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-21suggested: NoneSoftware and Algorithms Sentences Resources 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 Prismsuggested: (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.
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