Evaluation of heating and chemical protocols for inactivating SARS-CoV-2

  1. Boris Pastorino
  2. Franck Touret
  3. Magali Gilles
  4. Xavier de Lamballerie
  5. Remi N. Charrel

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Abstract

Clinical samples collected in COVID-19 patients are commonly manipulated in BSL-2 laboratories for diagnostic purpose. We used the French norm NF-EN-14476+A2 derived from the European standard EN-14885. To avoid the risk of exposure of laboratory workers, we showed that Triton-X100 must be added to guanidinium thiocyanate-lysis buffers to obtain a 6-log reduction of infectious virus. Although heating protocol consisting of 92°C-15min was more effective rather than 56°C-30min and 60°C-60min to achieve 6-log reduction, it is not amenable for molecular detection on respiratory specimens because of important decrease of detectable RNA copies in the treated sample vs untreated sample. The 56°C-30min and 60°C-60min should be used for inactivation of serum / plasma samples for serology because of the 5log10 reduction of infectivity and low viral loads in blood specimens.

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  1. ScreenIT

    SciScore for 10.1101/2020.04.11.036855: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    SARS-CoV-2 titration: SARS-CoV-2 was first propagated and titrated on Vero-E6 cells.
    Vero-E6
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Cell line: African green monkey kidney cells (Vero-E6; ATCC#CRL-1586) were grown at 37°C in 5% CO2 with 1% Penicillin/Streptomycin (PS; 5000U.mL-1 and 5000µg.mL-1; Life Technologies) and supplemented with 1% non-essential amino acids (Life Technologies) in Minimal Essential Medium (Life Technologies) with 5% FBS.
    Vero-E6; ATCC#CRL-1586
    suggested: None

    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).

    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.

  2. Version published to 10.1101/2020.04.11.036855v2 on bioRxiv
  3. Version published to 10.1101/2020.04.11.036855v1 on bioRxiv