Performance of a flow cytometry-based immunoassay for detection of antibodies binding to SARS-CoV-2 spike protein

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1. Version published to 10.1038/s41598-021-04565-1

   Jan 12, 2022
2. 

   ScreenIT

   Apr 11, 2021

   SciScore for 10.1101/2021.04.06.21254995: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: The study was approved by the Ethics Committee of Hospital Clínico Universitario INCLIVA.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   IgG or IgA antibodies bound to S proteins were identified by comparing the median fluorescence intensity (MFI) of the S-Jurkat and the 0-Jurkat cells in each sample.	IgA suggested: None
   Samples were considered positive for IgG or IgA when the normalized difference was ≥ 1. Commercially-available chemiluminescent SARS-CoV-2 S assays: Roche Elecsys® Anti-SARS-CoV-2 S (Roche …

   SciScore for 10.1101/2021.04.06.21254995: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: The study was approved by the Ethics Committee of Hospital Clínico Universitario INCLIVA.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   IgG or IgA antibodies bound to S proteins were identified by comparing the median fluorescence intensity (MFI) of the S-Jurkat and the 0-Jurkat cells in each sample.	IgA suggested: None
   Samples were considered positive for IgG or IgA when the normalized difference was ≥ 1. Commercially-available chemiluminescent SARS-CoV-2 S assays: Roche Elecsys® Anti-SARS-CoV-2 S (Roche Diagnostics, Pleasanton, CA, USA), an electrochemiluminescence sandwich immunoassay (ECLIA) that quantifies total (IgG and IgM) antibodies directed against RBD, was run on cobas® e601 modular analyzer (Roche Diagnostics, Rotkreuz, Switzerland).	total (IgG suggested: None IgM suggested: None
   The assay is calibrated with the first WHO International Standard and Reference Panel for anti-SARS-CoV-2 antibody (15).	anti-SARS-CoV-2 suggested: None
   LIAISON® SARS-CoV-2 TrimericS IgG assay (Diasorin S.p.a, Saluggia, Italy), run on a DiaSorin LIAISON platform (DiaSorin, Stillwater, USA), measured IgG antibodies against a trimeric S-protein antigen.	a trimeric S-protein antigen. suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Non-transfected Jurkat cells (0-Jutkat) were used as controls.	Jurkat suggested: None
   For each individual assay, a mixture of 50,000 0-Jurkat and 150,000 S-Jurkat cells was made in a single tube.	S-Jurkat suggested: None
   We established the difference between S-Jurkat and 0-Jurkat cells using the normalized MFI-ratio between EGFR and both antibody isotypes (IgG MFI-ratio and IgA MFI-ratio respectively).	0-Jurkat suggested: None
   Software and Algorithms
   Sentences	Resources
   The analyses were performed using SPSS version 20.0 (SPSS, Chicago, IL, USA).	SPSS suggested: (SPSS, RRID:SCR_002865)

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   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).

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   Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:

   In our view, the main limitations of the current study are the relative small number of specimens included in the evaluation panel and that discrepancies across results returned by the evaluated immunoassays were not resolved by performing antibody neutralization assays, the gold standard for serological diagnosis of SARS-CoV-2 infection (6). In summary, herein we have shown that a FCI using Jurkat T cells expressing the SARS-CoV-2 native S protein for detection of IgG and IgA-specific antibodies is highly specific and seemingly provides increased clinical sensitivity for diagnosis of SARS-CoV-2 infection when compared to two new-generation immunoassays targeting either the S protein in its trimeric conformation (Diasorin CLIA) or RBD (Roche ECLIA). The assay is easy to perform and standardize; the need for a flow cytometer should not be viewed as a disadvantage compared to high-throughput CLIA assays, as this platform is widely available at immunology and hematology departments in hospitals of all sizes. Further studies evaluating the performance of FCI for documenting seroconversion in vaccinated people are underway.

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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

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

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   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.

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3. 

   Version published to 10.1101/2021.04.06.21254995v1 on medRxiv

   Apr 9, 2021