Propagation, Inactivation, and Safety Testing of SARS-CoV-2
Propagation, Inactivation, and Safety Testing of SARS-CoV-2
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DOI：
10.3390/v12060622
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发表时间：
2020-06-01
期刊：
影响因子：
4.7
通讯作者：
Basler, Christopher F.
中科院分区：
文献类型：
--
作者：
Jureka, Alexander S.;Silvas, Jesus A.;Basler, Christopher F.
2019年底，一种新型冠状病毒，严重急性呼吸综合征冠状病毒2（SARS-CoV-2）在中国湖北省省会武汉市出现。从那时起，SARS-CoV-2已经造成了全球大流行，导致超过400万人感染和超过25万人死亡。这场大流行引发了与SARS-CoV-2及其引起的疾病COVID-19相关的广泛研究。对这种新病毒的研究将通过提供明确描述和有效的程序来促进，这些程序使感染性病毒的繁殖和定量成为可能。由于建议在生物安全等级3下进行病毒工作，因此还需要有效地鉴定病毒的有效方法，以便能够安全地研究受感染细胞的RNA、DNA和蛋白质。在这里，我们报告的方法用于在多种细胞系中生长SARS-CoV-2，并通过使用琼脂糖或微晶纤维素作为覆盖物的空斑试验以及SARS-CoV-2特异性病灶形成试验来测量病毒感染性。我们还证明了TRIzol，10%中性缓冲福尔马林，β丙内酯和热的有效灭活。
In late 2019, a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, the capital of the Chinese province Hubei. Since then, SARS-CoV-2 has been responsible for a worldwide pandemic resulting in over 4 million infections and over 250,000 deaths. The pandemic has instigated widespread research related to SARS-CoV-2 and the disease that it causes, COVID-19. Research into this new virus will be facilitated by the availability of clearly described and effective procedures that enable the propagation and quantification of infectious virus. As work with the virus is recommended to be performed at biosafety level 3, validated methods to effectively inactivate the virus to enable the safe study of RNA, DNA, and protein from infected cells are also needed. Here, we report methods used to grow SARS-CoV-2 in multiple cell lines and to measure virus infectivity by plaque assay using either agarose or microcrystalline cellulose as an overlay as well as a SARS-CoV-2 specific focus forming assay. We also demonstrate effective inactivation by TRIzol, 10% neutral buffered formalin, beta propiolactone, and heat.