An enzyme-based immunodetection assay to quantify SARS-CoV-2 infection

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1. Version published to 10.1016/j.antiviral.2020.104882

   Sep 1, 2020
2. 

   ScreenIT

   Jun 17, 2020

   SciScore for 10.1101/2020.06.14.150862: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   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
   After one hour, the cells were washed three times with washing buffer (0.3% (v/v) Tween 20 in PBS) before a secondary anti-mouse or anti-rabbit antibody conjugated with HRP was added (1:10,000, 1:15,000, 1:20,000 or 1:30,000) and incubated for 1 h at 37°C.	anti-mouse suggested: None anti-rabbit suggested: None
   Cells were then washed with PBS and stained with 1:5,000 diluted mouse anti-SARS-CoV-2 S protein antibody 1A9 (Biozol GTX-GTX632604) in antibody buffer …

   SciScore for 10.1101/2020.06.14.150862: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   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
   After one hour, the cells were washed three times with washing buffer (0.3% (v/v) Tween 20 in PBS) before a secondary anti-mouse or anti-rabbit antibody conjugated with HRP was added (1:10,000, 1:15,000, 1:20,000 or 1:30,000) and incubated for 1 h at 37°C.	anti-mouse suggested: None anti-rabbit suggested: None
   Cells were then washed with PBS and stained with 1:5,000 diluted mouse anti-SARS-CoV-2 S protein antibody 1A9 (Biozol GTX-GTX632604) in antibody buffer (PBS containing 10% (v/v) FCS and 0.3% (v/v) Tween 20) at 37°C.	anti-SARS-CoV-2 S protein suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Calu-3 (human epithelial lung adenocarcinoma) cells were cultured in Minimum Essential Medium Eagle (MEM, Sigma #M4655) supplemented with 10% FCS, 100 units/ml penicillin, 100 μg/ml streptomycin, 1 mM sodium pyruvate, and 1x non-essential amino acids.	Calu-3 suggested: None
   Virus isolation from patient samples: To isolate SARS-CoV-2 from patient samples, 50,000 Vero E6 cells were seeded in 24-well plates in 500 μl medium incubated over night at 37°C.	Vero E6 suggested: None
   To this end, 6,000 Vero E6 or 10,000 Caco-2 cells were seeded per well in 96 flat bottom well plates in 100 μl medium and incubated over night before 62 μl fresh medium was added.	Caco-2 suggested: None
   Software and Algorithms
   Sentences	Resources
   Statistical analysis: The determination of the inhibitory concentration 50 (IC50) or inhibitory titer 50 by four-parametric nonlinear regression and one-way ANOVA followed by Bonferroni’s multiple comparison test (ns not significant, * P < 0.01, ** P < 0.001, *** P < 0.0001) were performed using GraphPad Prism version 8.2.1 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798) GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)

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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: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

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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 10.1101/2020.06.14.150862v1 on bioRxiv

   Jun 15, 2020