tetano
Editor, Senior Moderator
Viruses
. 2021 Mar 26;13(4):562.
doi: 10.3390/v13040562.
Effect of Inactivation Methods on SARS-CoV-2 Virion Protein and Structure
Emma K Loveday[SUP] 1 [/SUP], Kyle S Hain[SUP] 2 [/SUP], Irina Kochetkova[SUP] 2 [/SUP], Jodi F Hedges[SUP] 2 [/SUP], Amanda Robison[SUP] 2 [/SUP], Deann T Snyder[SUP] 2 [/SUP], Susan K Brumfield[SUP] 3 [/SUP], Mark J Young[SUP] 3 [/SUP], Mark A Jutila[SUP] 2 [/SUP], Connie B Chang[SUP] 1 [/SUP], Matthew P Taylor[SUP] 2 [/SUP]
Affiliations
Abstract
The risk posed by Severe Acute Respiratory Syndrome Coronavirus -2 (SARS-CoV-2) dictates that live-virus research is conducted in a biosafety level 3 (BSL3) facility. Working with SARS-CoV-2 at lower biosafety levels can expedite research yet requires the virus to be fully inactivated. In this study, we validated and compared two protocols for inactivating SARS-CoV-2: heat treatment and ultraviolet irradiation. The two methods were optimized to render the virus completely incapable of infection while limiting the destructive effects of inactivation. We observed that 15 min of incubation at 65 ?C completely inactivates high titer viral stocks. Complete inactivation was also achieved with minimal amounts of UV power (70,000 ?J/cm[SUP]2[/SUP]), which is 100-fold less power than comparable studies. Once validated, the two methods were then compared for viral RNA quantification, virion purification, and antibody detection assays. We observed that UV irradiation resulted in a 2-log reduction of detectable genomes compared to heat inactivation. Protein yield following virion enrichment was equivalent for all inactivation conditions, but the quality of resulting viral proteins and virions were differentially impacted depending on inactivation method and time. Here, we outline the strengths and weaknesses of each method so that investigators might choose the one which best meets their research goals.
Keywords: ELISA; SARS-CoV-2; UV inactivation; electron microscopy; heat inactivation; western blot.
. 2021 Mar 26;13(4):562.
doi: 10.3390/v13040562.
Effect of Inactivation Methods on SARS-CoV-2 Virion Protein and Structure
Emma K Loveday[SUP] 1 [/SUP], Kyle S Hain[SUP] 2 [/SUP], Irina Kochetkova[SUP] 2 [/SUP], Jodi F Hedges[SUP] 2 [/SUP], Amanda Robison[SUP] 2 [/SUP], Deann T Snyder[SUP] 2 [/SUP], Susan K Brumfield[SUP] 3 [/SUP], Mark J Young[SUP] 3 [/SUP], Mark A Jutila[SUP] 2 [/SUP], Connie B Chang[SUP] 1 [/SUP], Matthew P Taylor[SUP] 2 [/SUP]
Affiliations
- PMID: 33810401
- DOI: 10.3390/v13040562
Abstract
The risk posed by Severe Acute Respiratory Syndrome Coronavirus -2 (SARS-CoV-2) dictates that live-virus research is conducted in a biosafety level 3 (BSL3) facility. Working with SARS-CoV-2 at lower biosafety levels can expedite research yet requires the virus to be fully inactivated. In this study, we validated and compared two protocols for inactivating SARS-CoV-2: heat treatment and ultraviolet irradiation. The two methods were optimized to render the virus completely incapable of infection while limiting the destructive effects of inactivation. We observed that 15 min of incubation at 65 ?C completely inactivates high titer viral stocks. Complete inactivation was also achieved with minimal amounts of UV power (70,000 ?J/cm[SUP]2[/SUP]), which is 100-fold less power than comparable studies. Once validated, the two methods were then compared for viral RNA quantification, virion purification, and antibody detection assays. We observed that UV irradiation resulted in a 2-log reduction of detectable genomes compared to heat inactivation. Protein yield following virion enrichment was equivalent for all inactivation conditions, but the quality of resulting viral proteins and virions were differentially impacted depending on inactivation method and time. Here, we outline the strengths and weaknesses of each method so that investigators might choose the one which best meets their research goals.
Keywords: ELISA; SARS-CoV-2; UV inactivation; electron microscopy; heat inactivation; western blot.