SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup.
SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup.
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DOI：
10.1002/cpmc.100
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发表时间：
2020-06-01
影响因子：
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通讯作者：
Krammer, Florian
中科院分区：
文献类型：
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作者：
Stadlbauer, Daniel;Amanat, Fatima;Krammer, Florian
2019年末，中国发现非典型肺炎病例。病原体很快被确定为贝塔冠状病毒(命名为SARS-CoV-2)，自那以来已导致大流行。已经建立了几种允许特定检测病毒核酸的方法，但这些方法只允许在短时间内检测病毒，通常是在急性感染期间。迫切需要进行血清学检测，以了解对病毒产生的抗体反应，并确定可能对再次感染具有免疫力的个人。在这里，我们描述了一种来自SARS-CoV-2刺突蛋白的抗原表达的详细方案，该方案可以作为免疫学分析的底物，以及一种两阶段血清学酶联免疫吸附试验(ELISA)。这些检测方法可用于研究研究和临床实验室的检测。©2020作者。基本方案1：哺乳动物细胞转染和蛋白质纯化基本方案2：用于高通量筛选与SARS-CoV-2刺突蛋白结合的抗体的人血清样本的两阶段ELISA法。
In late 2019, cases of atypical pneumonia were detected in China. The etiological agent was quickly identified as a betacoronavirus (named SARS-CoV-2), which has since caused a pandemic. Several methods allowing for the specific detection of viral nucleic acids have been established, but these only allow detection of the virus during a short period of time, generally during acute infection. Serological assays are urgently needed to conduct serosurveys, to understand the antibody responses mounted in response to the virus, and to identify individuals who are potentially immune to re-infection. Here we describe a detailed protocol for expression of antigens derived from the spike protein of SARS-CoV-2 that can serve as a substrate for immunological assays, as well as a two-stage serological enzyme-linked immunosorbent assay (ELISA). These assays can be used for research studies and for testing in clinical laboratories. © 2020 The Authors. Basic Protocol 1: Mammalian cell transfection and protein purification Basic Protocol 2: A two-stage ELISA for high-throughput screening of human serum samples for antibodies binding to the spike protein of SARS-CoV-2.