Evaluation of stability and inactivation methods of SARS-CoV-2 in context of laboratory settings

  • The novel coronavirus SARS-CoV-2 is the causative agent of the acute respiratory disease COVID-19, which has become a global concern due to its rapid spread. Laboratory work with SARS-CoV-2 in a laboratory setting was rated to biosafety level 3 (BSL-3) biocontainment level. However, certain research applications in particular in molecular biology require incomplete denaturation of the proteins, which might cause safety issues handling contaminated samples. In this study, we evaluated lysis buffers that are commonly used in molecular biological laboratories for their ability to inactivate SARS-CoV-2. In addition, viral stability in cell culture media at 4 °C and on display glass and plastic surfaces used in laboratory environment was analyzed. Furthermore, we evaluated chemical and non-chemical inactivation methods including heat inactivation, UV-C light, addition of ethanol, acetone-methanol, and PFA, which might be used as a subsequent inactivation step in the case of insufficient inactivation. We infected susceptible Caco-2 and Vero cells with pre-treated SARS-CoV-2 and determined the tissue culture infection dose 50 (TCID50) using crystal violet staining and microscopy. In addition, lysates of infected cells and virus containing supernatant were subjected to RT-qPCR analysis. We have found that guanidine thiocyanate and most of the tested detergent containing lysis buffers were effective in inactivation of SARS-CoV-2, however, the M-PER lysis buffer containing a proprietary detergent failed to inactivate the virus. In conclusion, careful evaluation of the used inactivation methods is required especially for non-denaturing buffers. Additional inactivation steps might be necessary before removal of lysed viral samples from BSL-3.
Author:Marek WideraORCiDGND, Sandra WesthausGND, Holger RabenauORCiDGND, Sebastian HoehlORCiDGND, Denisa BojkovaORCiDGND, Jindrich CinatlORCiDGND, Sandra CiesekORCiDGND
Parent Title (English):Medical microbiology and immunology
Place of publication:Berlin ; Heidelberg
Document Type:Article
Date of Publication (online):2021/07/01
Date of first Publication:2021/07/01
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2022/05/10
Tag:Bio safety; COVID-19; Corona virus; Inactivation; SARS-CoV-2; Stability
Page Number:10
First Page:235
Last Page:244
Open Access funding enabled and organized by Projekt DEAL. The authors have received research funding from Hönle AG, Germany. We thank the numerous donations and the support of SARS-CoV-2 research. MW and SC were supported by the Goethe-Corona-Fond of the Goethe University & University Hospital Frankfurt. M.W. was supported by the Deutsche Forschungsgemeinschaft (DFG, WI 5086/1–1).
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Licence (German):License LogoCreative Commons - Namensnennung 4.0