A serological assay to detect SARS-CoV-2 seroconversion in humans
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SciScore for 10.1101/2020.03.17.20037713: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: All participants provided written informed consent prior to the study.
IRB: The studies were approved by the Alfred Hospital (ID #280/14) and University of Melbourne (ID #1442952.1, 1955465.2) Human Research Ethics Committees, and under research permit for project TYH2018322 of Helsinki University Hospital Laboratory.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 Next, a 1:3000 dilution of goat anti-human IgG-horseradish peroxidase (HRP) conjugated secondary antibody (ThermoFisher Scientific) was … SciScore for 10.1101/2020.03.17.20037713: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: All participants provided written informed consent prior to the study.
IRB: The studies were approved by the Alfred Hospital (ID #280/14) and University of Melbourne (ID #1442952.1, 1955465.2) Human Research Ethics Committees, and under research permit for project TYH2018322 of Helsinki University Hospital Laboratory.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 Next, a 1:3000 dilution of goat anti-human IgG-horseradish peroxidase (HRP) conjugated secondary antibody (ThermoFisher Scientific) was prepared in 0.1% TPBS and 100 ul of this secondary antibody was added to each well for 1 hour. anti-human IgG-horseradishsuggested: NoneThese antibodies include anti-human IgA (α-chain-specific) HRP antibody (Sigma A0295) (1:3,000), anti-human IgM (μ-chain-specific) HRP antibody (Sigma A6907) (1:3,000), anti-human IgG1 Fc-HRP (Southern Biotech 9054-05) (1:3,000), anti-human IgG2 Fc-HRP (Southern Biotech #9060-05) (1:3,000), anti-human IgG3hinge-HRP (Southern Biotech 9210-05) (1:3,000), and anti-human IgG4 Fc-HRP (Southern Biotech 9200-05). anti-human IgAsuggested: (Sigma-Aldrich Cat# A0295, RRID:AB_257876)α-chain-specific) HRPsuggested: Noneanti-human IgMsuggested: (Sigma-Aldrich Cat# A6907, RRID:AB_258318)HRPsuggested: Noneanti-human IgG1suggested: (SouthernBiotech Cat# 9054-05, RRID:AB_2796627)anti-human IgG2suggested: (SouthernBiotech Cat# 9060-05, RRID:AB_2796633)anti-human IgG3hinge-HRPsuggested: Noneanti-human IgG4suggested: (SouthernBiotech Cat# 9200-05, RRID:AB_2796691)After blocking, 100 μL of 1C7 (anti-SARS NP antibody generated in house) at a dilution of 1:1000 was added to all wells and the plates were allowed to incubate for 1 hr at room temperature. anti-SARS NPsuggested: (Fitzgerald Industries International Cat# 20R-2831, RRID:AB_11196425)Experimental Models: Cell Lines Sentences Resources S1 proteins of NL63 and 229E were obtained from Sino Biologics (produced in 293HEK cells, hexa-histidine tagged). 293HEKsuggested: NonecDMEM was removed from Vero.E6 cells and 120 μL of the virus-serum mixture was added to the cells and the cells were incubated at 37°C for 1 hr. Vero.E6suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)Software and Algorithms Sentences Resources Recombinant proteins: The mammalian cell codon optimized nucleotide sequence coding for the spike protein of SARS-CoV-2 isolate (GenBank: MN908947.3) was synthesized commercially (GeneWiz). GeneWizsuggested: (GENEWIZ, RRID:SCR_003177)Several of the expression plasmids and proteins have also been submitted to BEI Resources and can be requested from their web page for free (https://www.beiresources.org/). https://www.beiresources.org/suggested: (BEI Resource Repository, RRID:SCR_013698)Data were analyzed using Prism 7 (Graphpad). Prismsuggested: (PRISM, RRID:SCR_005375)Graphpadsuggested: (GraphPad, RRID:SCR_000306)The following day, heat inactivated serum samples (dilution of 1:10) were serially diluted 3-fold in 2X MEM (20% 10× minimal essential medium (Gibco), 4⍰mM L-glutamine, 0.2% of sodium bicarbonate [wt/vol; Gibco], 20⍰mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, Gibco), 200⍰U/ml penicillin–200⍰μ/ml streptomycin (Gibco), and 0.4% bovine serum albumin (MP Biomedical)). Gibco]suggested: NoneAnalyses were performed in GraphPad Prism. 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: We found the following clinical trial numbers in your paper:
Identifier Status Title NCT04264858 Not yet recruiting Treatment of Acute Severe 2019-nCoV Pneumonia With Immunoglo… 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.
- No protocol registration statement was detected.
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