Multiplexed, quantitative serological profiling of COVID-19 from blood by a point-of-care test

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1. Version published to 10.1126/sciadv.abg4901

   Jun 25, 2021
2. 

   ScreenIT

   Nov 8, 2020

   SciScore for 10.1101/2020.11.05.20226654: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: Patient samples: De-identified heat-inactivated EDTA plasma samples (57°C for 30 minutes) were accessed from the Duke COVID-19 ICU biorepository (Pro00101196, PI Bryan Kraft) via an exempted protocol approved by the Duke University Institutional review board (Pro00105331, PI Ashutosh Chilkoti).
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Briefly, eligible patients included in the repository were men and women ages 18 years and above that were admitted to an adult ICU at Duke University Hospital with SARS-CoV-2 infection confirmed by PCR testing.
   Cell Line Authentication	not detected.

   Table 2: Resources

   …

   SciScore for 10.1101/2020.11.05.20226654: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: Patient samples: De-identified heat-inactivated EDTA plasma samples (57°C for 30 minutes) were accessed from the Duke COVID-19 ICU biorepository (Pro00101196, PI Bryan Kraft) via an exempted protocol approved by the Duke University Institutional review board (Pro00105331, PI Ashutosh Chilkoti).
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Briefly, eligible patients included in the repository were men and women ages 18 years and above that were admitted to an adult ICU at Duke University Hospital with SARS-CoV-2 infection confirmed by PCR testing.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   For DA-D4 assays that also detected IP-10, an additional column of five spots of capture antibody (R&D systems, cat# MAB266) was included and anti-IP-10 detection antibody (R&D systems, cat# AF-266) was included in the detection cocktail for the open format chips.	anti-IP-10 suggested: None
   To validate the analytical performance of the test, dose-response curves were generated using antibodies targeting SARS-CoV-2 antigens (Sino Biological, cat#: 40143-MM05, 40150-D001, and 40150-D004) spiked into undiluted pooled human serum.	antibodies targeting SARS-CoV-2 antigens suggested: (Sino Biological Cat# 40150-D001-H, RRID:AB_2857930)
   Experimental Models: Cell Lines
   Sentences	Resources
   These dilutions are transferred to a 96 well plate containing 2×104 Vero E6 cells per well.	Vero E6 suggested: RRID:CVCL_XD71)
   Software and Algorithms
   Sentences	Resources
   Once connected to the power source, the D4Scope automatically runs our custom imaging Python program.	Python suggested: (IPython, RRID:SCR_001658)
   For this study we manually analyzed the resulting fluorescence intensity using Genepix Analysis software.	Genepix Analysis suggested: None
   The assay was performed using a Beckman Coulter CytoFLEX flow cytometer and data processing was performed using BioLegend’s Bio-Bits cloud-based software platform.	BioLegend’s suggested: None
   Statistical analysis: Statistical analysis was performed using GraphPad Prism version 8.4.1 (GraphPad Software, Inc).	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: We detected the following sentences addressing limitations in the study:

   This limitation could be addressed by testing individual samples on separate microfluidic chips at various dilutions, which would effectively increase the dynamic range of our assay and yield more precise quantitative titer. Additionally, because of the double antigen design of our assay, we are also not able to discriminate between specific antibody subclasses or isotypes, which has been shown to be important for other diseases. Despite these limitations, we believe our assay is well poised to complement existing diagnostic solutions once additional validation studies encompassing larger patient cohorts are completed. In summary, we have developed a COVID-19 serological assay that merges the benefits of LFAs and ELISAs. We used this test to simultaneously measure the antibody levels for multiple viral antigens and a potential prognostic biomarker directly from plasma and whole blood. For COVID-19 management, our platform may be useful to better understand patient antibody responses, provide actionable intelligence to physicians to guide interventions for hospitalized patients at the point-of-care, to assess vaccine efficacy, and to perform epidemiological studies. Further, our platform is broadly applicable to other diseases where sensitive and quantitative antibody and or protein detection is desirable in settings without access to a centralized laboratory. Overall, we believe that our platform is a promising approach to democratize access to laboratory quality tests, by en...

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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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   Read the original source
3. 

   Version published to 10.1101/2020.11.05.20226654v1 on medRxiv

   Nov 7, 2020