Multiplex PCR Assays for Identifying all Major Severe Acute Respiratory Syndrome Coronavirus 2 Variants

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Abstract

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  1. SciScore for 10.1101/2021.10.28.21263107: (What is this?)

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

    Table 1: Rigor

    Ethicsnot detected.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line AuthenticationAuthentication: qRT-PCR screening was performed using 2 µL of purified RNA, TaqPath 1-step Multiplex Master Mix (No ROX) and an in-house validated multiplex assay.

    Table 2: Resources

    Software and Algorithms
    SentencesResources
    Synthetic RNA targets: RNA standards of the SARS-CoV-2 whole genome and variant spike proteins were provided by Exact Diagnostics through Bio-Rad Laboratories, Hercules, CA (catalog numbers COV019, COV019CE, COVA, COVB, COVE, COVG).
    Bio-Rad Laboratories
    suggested: (Bio-Rad Laboratories, RRID:SCR_008426)
    Primer Design: Primers were designed by using the primer3 server13.
    primer3
    suggested: (Primer3, RRID:SCR_003139)
    The desired fractions were then ethanol precipitated and resuspended in water, and their concentration was determined by NanoDrop spectrophotometer.
    NanoDrop
    suggested: None
    Multiplex qRT-PCR with patient samples: Patient samples were tested in a similar manner to how we tested the assay’s sensitivity and specificity. 5 µL of each sample was combined with 15 µL each completed master mix using TaqPath and run as mentioned before.
    TaqPath
    suggested: None

    Results from OddPub: Thank you for sharing your 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.

    Results from scite Reference Check: We found no unreliable references.


    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.