A simple protein-based surrogate neutralization assay for SARS-CoV-2
A simple protein-based surrogate neutralization assay for SARS-CoV-2
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DOI：
10.1172/jci.insight.142362
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发表时间：
2020-10-02
期刊：
影响因子：
8
通讯作者：
Gingras, Anne-Claude
中科院分区：
文献类型：
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作者：
Abe, Kento T.;Li, Zhijie;Gingras, Anne-Claude
大多数感染严重急性呼吸综合征冠状病毒2（SARS - CoV - 2）的患者在感染后的几周内会对病毒产生体液免疫反应，但这种反应的持续时间以及它与临床结果的相关性尚未完全明确。尤其重要的是确定感染的免疫相关因素，这将为治疗方法、疫苗接种策略和恢复期血浆疗法的公共卫生决策提供支持。虽然已经开发出基于酶联免疫吸附测定（ELISA）的方法来检测和定量患者样本中针对SARS - CoV - 2的抗体，但中和抗体的检测通常需要要求更高的基于细胞的病毒检测方法。在此，我们介绍一种安全且高效的基于蛋白质的检测方法，用于检测血清和血浆中阻断SARS - CoV - 2刺突蛋白受体结合域（RBD）与其受体血管紧张素转换酶2（ACE2）相互作用的抗体。该检测方法可作为一种替代中和检测方法，并且与用于检测针对RBD抗体的ELISA在同一平台上同时进行，从而能够进行直接比较。我们的检测方法所获得的结果与两种基于病毒的检测方法相关，一种是使用活SARS - CoV - 2病毒的空斑减少中和试验（PRNT），另一种是基于刺突假型病毒载体的检测方法。
Most of the patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mount a humoral immune response to the virus within a few weeks of infection, but the duration of this response and how it correlates with clinical outcomes has not been completely characterized. Of particular importance is the identification of immune correlates of infection that would support public health decision-making on treatment approaches, vaccination strategies, and convalescent plasma therapy. While ELISA-based assays to detect and quantitate antibodies to SARS-CoV-2 in patient samples have been developed, the detection of neutralizing antibodies typically requires more demanding cell-based viral assays. Here, we present a safe and efficient protein-based assay for the detection of serum and plasma antibodies that block the interaction of the SARS-CoV-2 spike protein receptor binding domain (RBD) with its receptor, angiotensin-converting enzyme 2 (ACE2). The assay serves as a surrogate neutralization assay and is performed on the same platform and in parallel with an ELISA for the detection of antibodies against the RBD, enabling a direct comparison. The results obtained with our assay correlate with those of 2 viral-based assays, a plaque reduction neutralization test (PRNT) that uses live SARS-CoV-2 virus and a spike pseudotyped viral vector-based assay.