SARS-CoV-2 evolution during treatment of chronic infection

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Abstract

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  1. SciScore for 10.1101/2020.12.05.20241927: (What is this?)

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

    Table 1: Rigor

    NIH rigor criteria are not applicable to paper type.

    Table 2: Resources

    Antibodies
    SentencesResources
    The SARSCOV-2 Spike proteins were visualized by a ChemiDoc® MP imaging system (Biorad) using anti-Spike S2 (Invitrogen) and anti-p24 Gag antibodies.
    anti-Spike S2
    suggested: (Imported from the IEDB Cat# S2, RRID:AB_2833224)
    anti-p24 Gag antibodies.
    suggested: None
    Testing of convalescent plasma for antibody titres: The Anti-SARS-CoV-2 ELISA (IgG) assay used to test CP for antibody titres was Euroimmun Medizinische Labordiagnostika AG.
    Anti-SARS-CoV-2 ELISA (IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    This indirect ELISA based assay uses a recombinant structural spike 1 (S1) protein of SARS-CoV-2 expressed in the human cell line HEK 29 for the detection of SARS-CoV2 IgG.
    HEK 29
    suggested: None
    Virus neutralisation assays were performed on 293T cell transiently transfected with ACE2 and TMPRSS2 using SARS-CoV-2 Spike pseudotyped virus expressing luciferase48.
    293T
    suggested: None
    mAb pseudotype neutralisation assay: Virus neutralisation assays were performed on HeLa cells stably expressing ACE2 and using SARS-CoV-2 Spike pseudotyped virus expressing luciferase as previously described49.
    HeLa
    suggested: None
    Then, freshly trypsinized HeLa ACE2-expressing cells were added to each well.
    HeLa ACE2-expressing
    suggested: None
    Software and Algorithms
    SentencesResources
    For short-read sequencing, FASTQs were downloaded, poor-quality reads were identified and removed, and both Illumina and PHiX adapters were removed using TrimGalore v0.6.6 28.
    TrimGalore
    suggested: None
    BAM files were then sorted and indexed with samtools v1.11 and PCR optical duplicates removed using Picard (http://broadinstitute.github.io/picard).
    samtools
    suggested: (SAMTOOLS, RRID:SCR_002105)
    Picard
    suggested: (Picard, RRID:SCR_006525)
    All sequences were aligned to the SARS-CoV-2 reference strain MN908947.3, using MAFFT v7.475 with automatic flavour selection34.
    MAFFT
    suggested: (MAFFT, RRID:SCR_011811)
    Maximum likelihood phylogenetic trees were produced using the above curated dataset using IQ-TREE v2.1.236.
    IQ-TREE
    suggested: (IQ-TREE, RRID:SCR_017254)
    All trees were visualised with Figtree v.
    Figtree
    suggested: (FigTree, RRID:SCR_008515)
    In-depth allele frequency variant calling: The SAMFIRE package39 was used to call allele frequency trajectories from BAM file data.
    SAMFIRE
    suggested: None
    The Mathematica software package was to conduct a regression analysis of pairwise sequence distances against time, leading to an estimate of a mean rate of within-host sequence evolution.
    Mathematica
    suggested: (Wolfram Mathematica, RRID:SCR_014448)
    To corroborate any findings, ClonalFrameML v1.12 42 was also used to infer recombination breakpoints.
    ClonalFrameML
    suggested: (Clonalframe, RRID:SCR_016060)
    Structural Viewing: The Pymol Molecular Graphics System v2.4.0 (https://github.com/schrodinger/pymol-open-source/releases) was used to map the location of the four spike mutations of interested onto a SARS-CoV-2 spike structure visualised by Wrobel et al (PDB: 6ZGE)43.
    Pymol
    suggested: (PyMOL, RRID:SCR_000305)
    IC50 values were calculated in GraphPad Prism.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: Thank you for sharing your code and data.


    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: No clinical trial numbers were referenced.


    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.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.