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 50 ) 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. Supplementary Information The online version contains supplementary material available at 10.1007/s00430-021-00716-3.
【저자키워드】 COVID-19, SARS-CoV-2, Corona virus, stability, inactivation, Bio safety, 【초록키워드】 Molecular biology, Infection, media, Proteins, virus, Novel coronavirus, Ethanol, Laboratory, Spread, RT-qPCR, Culture, lysis buffer, inactivation, heat inactivation, Microscopy, Cell culture, Research, molecular, Analysis, dose, Caco-2, acute respiratory disease, tissue culture, causative agent, biosafety level, level 3, culture media, guanidine, methanol, Crystal violet, supplementary material, tissue, infected cells, novel coronavirus SARS-CoV-2, removal, Vero cells, handling, staining, lysates, infected cell, denaturation, TCID, supernatant, Vero Cell, acetone, lysate, PFA, susceptible, effective, Biosafety Level 3, BSL-3, tested, analyzed, subsequent, addition, evaluated, required, lysed, viral sample, 【제목키워드】 Laboratory,