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Evaluation of stability and inactivation methods of SARS-CoV-2 in context of laboratory settings.
- Source :
-
Medical microbiology and immunology [Med Microbiol Immunol] 2021 Aug; Vol. 210 (4), pp. 235-244. Date of Electronic Publication: 2021 Jul 01. - Publication Year :
- 2021
-
Abstract
- 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 (TCID <subscript>50</subscript> ) 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.<br /> (© 2021. The Author(s).)
- Subjects :
- Animals
Caco-2 Cells
Cell Line
Chlorocebus aethiops
Containment of Biohazards
Humans
RNA, Viral
Real-Time Polymerase Chain Reaction
SARS-CoV-2 physiology
Specimen Handling methods
Time Factors
Vero Cells
Anti-Infective Agents pharmacology
COVID-19 prevention & control
COVID-19 virology
Guanidines pharmacology
SARS-CoV-2 drug effects
Thiocyanates pharmacology
Virus Inactivation
Subjects
Details
- Language :
- English
- ISSN :
- 1432-1831
- Volume :
- 210
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Medical microbiology and immunology
- Publication Type :
- Academic Journal
- Accession number :
- 34196781
- Full Text :
- https://doi.org/10.1007/s00430-021-00716-3