Ammonium Sulfate Addition Reduces the Need for Guanidinium Isothiocyanate in the Denaturing Transport Medium Used for SARS-COV-2 RNA Detection
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Abstract
Rapid identification of SARS-CoV-2 infected individuals through viral RNA detection followed by effective personal isolation remains the most effective way to prevent the spread of this virus. Large-scale RNA detection involves mass specimen collection and transportation. For biosafety reasons, denaturing viral transport medium has been extensively used during the pandemic. But the high concentrations of guanidinium isothiocyanate (GITC) in such media have raised issues around sufficient GITC supply and laboratory safety. Here, we tested whether supplementing media containing low concentrations of GITC with ammonium sulfate (AS) would affect the throat-swab detection of SARS-CoV-2 pseudovirus or a viral inactivation assay targeting both enveloped and non-enveloped viruses. Adding AS to the denaturing transport media reduced the need for high levels of GITC and improved SARS-COV-2 RNA detection without compromising virus inactivation.
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SciScore for 10.1101/2022.02.28.22271591: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources 2.1 Cells and viruses: Vero cells (ATCC, CCL-81) were cultured in Dulbecco’s modified Eagle’s medium (Gibco, NY, USA) supplemented with 10% (v/v) fetal calf serum. Verosuggested: NoneSoftware and Algorithms Sentences Resources FNV-2019-nCoV-abEN, >108 copies/mL) was purchased from Fubio Biological Technology Co., Ltd. (Hangzhou, China) Fubio Biologicalsuggested: NoneGraphs and statistical analyses (multiple t tests) were prepared with GraphPad Prism 8.0.2 (GraphPad … SciScore for 10.1101/2022.02.28.22271591: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources 2.1 Cells and viruses: Vero cells (ATCC, CCL-81) were cultured in Dulbecco’s modified Eagle’s medium (Gibco, NY, USA) supplemented with 10% (v/v) fetal calf serum. Verosuggested: NoneSoftware and Algorithms Sentences Resources FNV-2019-nCoV-abEN, >108 copies/mL) was purchased from Fubio Biological Technology Co., Ltd. (Hangzhou, China) Fubio Biologicalsuggested: NoneGraphs and statistical analyses (multiple t tests) were prepared with GraphPad Prism 8.0.2 (GraphPad Software, Inc, La Jolla, CA, United States). GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)GraphPadsuggested: (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: 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.
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