A rapid, point-of-care red blood cell agglutination assay detecting antibodies against SARS-CoV-2

Article activity feed

1. Version published to 10.1016/j.bbrc.2021.03.016

   May 1, 2021
2. 

   ScreenIT

   Jun 8, 2020

   SciScore for 10.1101/2020.05.13.094490: (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
   For testing patient serum, a dilution series of RBD-scFv was performed to test for optimal levels of protein to induce agglutination in presence of patient anti-RBD antibodies.	anti-RBD suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   (hyperactive piggyBac transposase) were co-transfected using Lipofectamine 3000 into HEK 293T cells.	HEK 293T suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
   Software and Algorithms
   Sentences	Resources
   One prep of 800 …

   SciScore for 10.1101/2020.05.13.094490: (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
   For testing patient serum, a dilution series of RBD-scFv was performed to test for optimal levels of protein to induce agglutination in presence of patient anti-RBD antibodies.	anti-RBD suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   (hyperactive piggyBac transposase) were co-transfected using Lipofectamine 3000 into HEK 293T cells.	HEK 293T suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
   Software and Algorithms
   Sentences	Resources
   One prep of 800 μL supernatant through one column of the kit yielded 102 μg/mL of RBD-scFv measured by NanoDrop™ 2000/2000c Spectrophotometers (ThermoFisher), which was used in subsequent experiments.	NanoDrop™ suggested: None ThermoFisher suggested: (ThermoFisher; SL 8; Centrifuge, RRID:SCR_020809)

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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:

   A key limitation of this assay is that it does not distinguish between IgG, IgA, or IgM against SARS-CoV-2, which may be desired in certain clinical scenarios. IgG subclasses can similarly not be distinguished. Also, the assay is highly dependent on the viral antigen deployed, such that clinicians should be aware if RBD antibody levels are negative, the patient could have still had COVID19. While the assay is simple and can be read with the naked eye, there is more subjectivity to it compared to interpreting lines on lateral flow assays or light detection in chemiluminescent ELISA’s. Depending on mixing technique and provider comfort, there could be variability, likely necessitating training and/or deployment of controls in order to ensure proper reading of the assay. A negative control test without fusion protein will be important to include during clinical implementation for patients with positive tests, given that rare patients may have IgM autoantibodies causing agglutination.31 Given the similarities of this test to currently used ABO typing32 and Monospot assays for EBV antibodies,33 we ultimately do not see this being a significant barrier. Lastly, the fusion protein stability is unknown, which is important for use in low-resource countries. Past studies with similar fusion proteins for hemagglutination assays found stability at least 30 days at 37°C and 6 months at 4°C with no loss of assay agglutination activity.14 Ultimately, similar investigations will be important...

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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 10.1101/2020.05.13.094490v1 on bioRxiv

   May 14, 2020