Analytical and clinical performances of a SARS-CoV-2 S-RBD IgG assay: comparison with neutralization titers

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1. Version published to 10.1515/cclm-2021-0313

   Apr 14, 2021
2. 

   ScreenIT

   Mar 15, 2021

   SciScore for 10.1101/2021.03.10.21253260: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: The study protocol (number 23307) was approved by the Ethics Committee of the University-Hospital, Padova. Consent: All the patients were informed of the study and voluntarily agreed to participate, providing a written consent.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	Considering a type I error α = 0.05, a power of 0.8 and with 249 positive and 249 negative subjects, a sensitivity (or specificity) of 0.95 can be considered significant with respect to values above or equal to 0.99 (null hypothesis).
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   2.5 Plaque …

   SciScore for 10.1101/2021.03.10.21253260: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: The study protocol (number 23307) was approved by the Ethics Committee of the University-Hospital, Padova. Consent: All the patients were informed of the study and voluntarily agreed to participate, providing a written consent.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	Considering a type I error α = 0.05, a power of 0.8 and with 249 positive and 249 negative subjects, a sensitivity (or specificity) of 0.95 can be considered significant with respect to values above or equal to 0.99 (null hypothesis).
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   2.5 Plaque reduction neutralization test (PRNT): A high-throughput PRNT method was used for the fast and accurate quantification of neutralizing antibodies in plasma samples collected from patients exposed to SARS-CoV-2, as described elsewhere (12).	SARS-CoV-2 suggested: None
   After 26 h of incubation and cells fixing, visualization of plaques was obtained with an immunocytochemical staining method using an anti-dsRNA monoclonal antibody (J2, 1:10,000; Scicons) for 1 hour, followed by 1-hour incubation with peroxidase-labeled goat anti-mouse antibodies (1:1000; DAKO) and a 7-minute incubation with the True Blue™ (KPL) peroxidase substrate.	anti-dsRNA suggested: (Millipore Cat# MABE1134, RRID:AB_2819101) J2 suggested: (US Biological Cat# U1000-87M, RRID:AB_2210756) anti-mouse suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   After 1 hour at 37 °C, fifty microliters of the virus–serum mixtures were added to confluent monolayers of Vero E6 cells, in 96-wells plates and incubated for 1 hour at 37 °C, in a 5% CO2 incubator.	Vero E6 suggested: None
   Software and Algorithms
   Sentences	Resources
   The GraphPad Prism version 9.1 for Windows (GraphPad Software, LLC) was employed to evaluate plaque reduction neutralization test results.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798) GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
   Stata v16.1 (Statacorp, Lakeway Drive, TX, USA) was used to evaluate the assays’ clinical performances.	Statacorp suggested: None

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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:

   This study presented several limitations. First, neutralizing antibodies were mainly tested in a well-defined cohort of family cluster, with sera collected at various time points and, therefore, should be confirmed in further studies; second, COVID-19 positive patients were selected retrospectively on the basis of available leftover samples, and third cross-reactivity with seasonal human coronaviruses was not assessed; therefore NPV and PPV could be overestimated. Another limitation of this study is that no longitudinal sera were analyzed and, therefore, we cannot exclude that some patients might have seroconverted at later time points. In conclusion, the data reported in this study showed that anti-SARS-CoV-2 S-RBD IgG assay achieves excellent analytical and clinical performances. Since specificity results were not 100%, the assay might present a limited number of false-positive results and this characteristic could be further confirmed in a more representative number of samples. However, the correlation with sera neutralization activity was very elevated, demonstrating that the dynamic range of the assay is expanded enough to capture all clinically significant NAb results. Finally, an appropriate threshold could be derived for selecting samples for COVID-19 convalescent plasma therapy.

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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 10.1101/2021.03.10.21253260v1 on medRxiv

   Mar 12, 2021