Reverse-Transcription Loop-Mediated Isothermal Amplification Has High Accuracy for Detecting Severe Acute Respiratory Syndrome Coronavirus 2 in Saliva and Nasopharyngeal/Oropharyngeal Swabs from Asymptomatic and Symptomatic Individuals

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Abstract

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  1. SciScore for 10.1101/2021.06.28.21259398: (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 Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    0% confluent Vero E6 cells and allowed to adsorb for 1 hour.
    Vero E6
    suggested: RRID:CVCL_XD71)
    Software and Algorithms
    SentencesResources
    SARS-CoV-2 Real-Time RT-qPCR: RNA was analysed using a range of different methods available at each site: CerTest VIASURE SARS-CoV-2 real time qPCR assay: Single step RT-qPCR against the ORF1ab region and N1 gene target of SARS-CoV-2 was carried out using the CerTest VIASURE SARS-CoV-2 real time PCR kit (CerTest Biotech SL, Zaragoza, Spain) according to manufacturer’s instructions for use (IFU) on either the ThermoFisher QuantStudio 5 or BioMolecular Systems MIC instruments, using 5 µl of extracted RNA per reaction.
    ThermoFisher QuantStudio
    suggested: (Primer Express Software, RRID:SCR_017376)
    Real-Time qPCR assay: Single step RT-qPCR against the E gene target of SARS-CoV-2 was carried out with the Corman et al. 3 primers using the AgPath-ID™ PCR kit (Thermofisher) according to manufacturer’s instructions for use (IFU) on an Aria qPCR Cycler (Agilent) and results analysed using the Agilent AriaMX 1.5 software, using 5 µl of extracted RNA per reaction.
    Agilent AriaMX
    suggested: (Agilent AriaMx Real-time PCR System, RRID:SCR_019469)
    Confusion matrices, sensitivity, specificity, sensitivity as a function of viral load calculations, and the production of scatter graphs showing the relationship between RT-LAMP results and CT were performed using Python 3.8.6.
    Python
    suggested: (IPython, RRID:SCR_001658)

    Results from OddPub: Thank you for sharing your data.


    Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:
    Incorporation of this inhibition control into the RT-LAMP assays would alleviate a potential limitation of the current assays and further support quality assurance for use within a clinical diagnostic setting. This study demonstrated high sensitivity and specificity for a novel sample preparation method used for SARS-CoV-2 Direct RT-LAMP, particularly in samples from which the individual would likely be considered infectious, highlighting the usefulness of saliva as a simple to collect, non-invasive sample type. The highly sensitive RNA RT-LAMP assay provides a rapid alternative with a reliance on differing reagents and equipment to RT-qPCR testing, thus providing additional diagnostic capacity and redundancy through diversity. Direct RT-LAMP may complement existing surveillance tools for SARS-CoV-2 testing including other point-of-care and laboratory-based diagnostics and is applicable to a variety of clinical scenarios, such as frequent, interval-based testing of asymptomatic individuals that may be missed when reliance is on symptomatic testing alone. However, care should be taken when considering frequency of testing, messaging around the role and interpretation of asymptomatic rapid tests, integration of data storage and access, and the challenges faced when scaling up surveillance to large populations. The role out of a new testing strategy can often throw up interesting and unexpected experiences. These collective experiences and lessons learnt from setting up an NHS a...

    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.