Humidity and Deposition Solution Play a Critical Role in Virus Inactivation by Heat Treatment of N95 Respirators
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Abstract
Shortages of personal protective equipment, including N95 respirators, during the coronavirus (CoV) disease 2019 (COVID-19) pandemic have highlighted the need to develop effective decontamination strategies for their reuse. This is particularly important in health care settings for reducing exposure to respiratory viruses, like severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes COVID-19. Although several treatment methods are available, a widely accessible strategy will be necessary to combat shortages on a global scale. We demonstrate that the combination of heat and humidity inactivates a range of RNA viruses, including both viral pathogens and common viral pathogen surrogates, after deposition on N95 respirators and achieves the necessary virus inactivation detailed by the U.S. Food and Drug Administration guidelines to validate N95 respirator decontamination technologies. We further demonstrate that depositing viruses onto surfaces when suspended in culture media can greatly enhance observed inactivation, adding caution to how heat and humidity treatment methods are validated.
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SciScore for 10.1101/2020.06.22.20137448: (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
Experimental Models: Cell Lines Sentences Resources IAV was enumerated in MDCK-SIAT1 cells by endpoint dilution using IAV titer media, which contained 1% BSA, but otherwise had the same components as IAV medium. MDCK-SIAT1suggested: ECACC Cat# 05071502, RRID:CVCL_Z936)Software and Algorithms Sentences Resources After one hour of incubation at room temperature with rocking, the virus suspension was removed from monolayers and an overlay of 1.6% agar mixed 1:1 with a 2x Minimum Essential … SciScore for 10.1101/2020.06.22.20137448: (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
Experimental Models: Cell Lines Sentences Resources IAV was enumerated in MDCK-SIAT1 cells by endpoint dilution using IAV titer media, which contained 1% BSA, but otherwise had the same components as IAV medium. MDCK-SIAT1suggested: ECACC Cat# 05071502, RRID:CVCL_Z936)Software and Algorithms Sentences Resources After one hour of incubation at room temperature with rocking, the virus suspension was removed from monolayers and an overlay of 1.6% agar mixed 1:1 with a 2x Minimum Essential Medium (MEM; Quality Biological, Cat. No. 115073101) containing 5% horse serum, 10 mM HEPES (Lonza, Cat. No. 17737E), 1X MEM Non-essential amino acids (Invitrogen, Cat. No. 11140050), 2% L-glutamine, and 2% penicillin streptomycin were applied. Quality Biologicalsuggested: NoneStatistical analyses: Unpaired t-tests were performed to determine differences in virus inactivation for different treatment conditions and viruses using GraphPad Prism 8 software. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)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: We detected the following sentences addressing limitations in the study:These limitations of the IAV and MHV experiments affect our ability to observe major trends in their relative susceptibilities. Nonetheless, the results from experiments with all four viruses suggest that bacteriophages are not always conservative surrogates for IAV and coronavirus inactivation through heat and humidity treatment. This brings to question the USFDA’s guidelines for using non-enveloped viruses as conservative surrogates for pathogenic viruses on N95 respirator decontamination. A review by Yang and Marr on virus survival in aerosols indicates that the presence of a lipid envelope is not solely responsible for virus susceptibility to inactivation; they suggest other virus characteristics, such as virus infection mechanisms and protein stability, are necessary to explain observed inactivation levels (51). To better account for these differences, a “cocktail” approach to assessing virus inactivation may be most suitable, using a wide range of surrogate viruses that have various characteristics in common with the viral pathogens of interest. Effects of deposition solution: Our results demonstrate that the deposition solution used to apply viruses to N95 respirators greatly impacts virus inactivation, both at elevated and ambient temperatures. At elevated temperatures, both MS2 and phi6 were inactivated much more when deposited in their culture media relative to PBS (Figures 2 and 3). At room temperature, only MS2 was inactivated to a greater extent when deposited in...
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.
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- No protocol registration statement was detected.
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