Growth, detection, quantification, and inactivation of SARS-CoV-2
Growth, detection, quantification, and inactivation of SARS-CoV-2
复制标题
DOI：
10.1016/j.virol.2020.05.015
复制
发表时间：
2020-09-01
期刊：
影响因子：
3.7
通讯作者：
Diamond, Michael S.
中科院分区：
文献类型：
--
作者：
Case, James Brett;Bailey, Adam L.;Diamond, Michael S.
严重急性呼吸综合征冠状病毒（SARS-CoV）-2是导致2019冠状病毒病（COVID-19）全球大流行的病原体。SARS-CoV-2与引起2003年SARS爆发的SARS-CoV密切相关。尽管许多试剂被开发用于研究SARS-CoV感染，但很少有适用于评价SARS-CoV-2感染和免疫。目前研究SARS-CoV-2的局限性包括很少有完全复制能力的野生型病毒的有效检测。我们已经制定了传播、量化和处理传染性SARS-CoV-2的方案。在这里，我们描述：（1）病毒储备液生成，（2）SARS-CoV-2 RNA的RTqPCR定量;（3）通过流式细胞术检测SARS-CoV-2抗原，（4）通过病灶形成和空斑试验定量感染性SARS-CoV-2;（5）病毒灭活的验证方案。总的来说，这些方法可以适用于各种实验设计，这将加速我们对SARS-CoV-2生物学的理解，并制定有效的COVID-19对策。
Severe acute respiratory syndrome coronavirus (SARS-CoV)-2 is the agent responsible for the coronavirus disease 2019 (COVID-19) global pandemic. SARS-CoV-2 is closely related to SARS-CoV, which caused the 2003 SARS outbreak. Although numerous reagents were developed to study SARS-CoV infections, few have been applicable to evaluating SARS-CoV-2 infection and immunity. Current limitations in studying SARS-CoV-2 include few validated assays with fully replication-competent wild-type virus. We have developed protocols to propagate, quantify, and work with infectious SARS-CoV-2. Here, we describe: (1) virus stock generation, (2) RTqPCR quantification of SARS-CoV-2 RNA; (3) detection of SARS-CoV-2 antigen by flow cytometry, (4) quantification of infectious SARS-CoV-2 by focus-forming and plaque assays; and (5) validated protocols for virus inactivation. Collectively, these methods can be adapted to a variety of experimental designs, which should accelerate our understanding of SARS-CoV-2 biology and the development of effective countermeasures against COVID-19.