Effect of Inactivation Methods on SARS-CoV-2 Virion Protein and Structure.
Effect of Inactivation Methods on SARS-CoV-2 Virion Protein and Structure.
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DOI：
10.3390/v13040562
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发表时间：
2021-03-26
期刊：
影响因子：
--
通讯作者：
Taylor MP
中科院分区：
文献类型：
--
作者：
Loveday EK;Hain KS;Kochetkova I;Hedges JF;Robison A;Snyder DT;Brumfield SK;Young MJ;Jutila MA;Chang CB;Taylor MP
严重急性呼吸综合征冠状病毒-2(SARS-CoV-2)带来的风险要求在生物安全3级(BSL3)设施中进行活病毒研究。在较低的生物安全水平下与SARS-CoV-2合作可以加快研究速度，但需要病毒完全灭活。在这项研究中，我们验证并比较了两种灭活SARS-CoV-2的方案：热处理和紫外线照射。这两种方法都经过了优化，使病毒完全不能感染，同时限制了灭活的破坏性影响。我们观察到，在65°C下孵育15分钟完全灭活高滴度病毒储存。在最小的紫外线功率(70,000微焦耳/厘米~2)下也实现了完全灭活，这比同类研究的功率低100倍。一旦得到验证，两种方法随后在病毒RNA定量、病毒粒子纯化和抗体检测分析方面进行了比较。我们观察到，与热灭活相比，紫外线照射导致可检测到的基因组减少了2个对数。病毒粒子浓缩后的蛋白质产量在所有灭活条件下都是相同的，但所产生的病毒蛋白和病毒粒子的质量受到不同灭活方法和时间的不同影响。在这里，我们概述了每种方法的优点和缺点，以便研究人员可以选择最符合他们的研究目标的方法。
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 µJ/cm2), 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.
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影响因子：
2.7
作者：
Cerutti H;Ricci V;Tesi G;Soldatini C;Castria M;Vaccaro MN;Tornesi S;Toppi S;Verdiani S;Brogi A
通讯作者：
Brogi A
影响因子：
3.1
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Darnell ME;Subbarao K;Feinstone SM;Taylor DR
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影响因子：
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Roberts ADG
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通讯作者：
Basler, Christopher F.
影响因子：
5.4
作者：
Neuman, Benjamin W.;Adair, Brian D.;Buchmeier, Michael J.
通讯作者：
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