SARS-CoV-2 Assays To Detect Functional Antibody Responses That Block ACE2 Recognition in Vaccinated Animals and Infected Patients.
SARS-CoV-2 Assays To Detect Functional Antibody Responses That Block ACE2 Recognition in Vaccinated Animals and Infected Patients.
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DOI：
10.1128/jcm.01533-20
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发表时间：
2020-10-21
影响因子：
9.4
通讯作者：
Kulp DW
中科院分区：
文献类型：
--
作者：
Walker SN;Chokkalingam N;Reuschel EL;Purwar M;Xu Z;Gary EN;Kim KY;Helble M;Schultheis K;Walters J;Ramos S;Muthumani K;Smith TRF;Broderick KE;Tebas P;Patel A;Weiner DB;Kulp DW
严重急性呼吸综合征冠状病毒2 （SARS-CoV-2）引起了COVID-19全球大流行，导致轻度至重度呼吸窘迫病例和大量死亡率。这种新型冠状病毒的全球爆发目前已在全球感染了2000万人，其中美国感染了500万例（2020年8月11日）。迫切需要开发诊断和研究工具，以确定感染和疫苗效力。严重急性呼吸综合征冠状病毒2 （SARS-CoV-2）引起了COVID-19全球大流行，导致轻度至重度呼吸窘迫病例和大量死亡率。这种新型冠状病毒的全球爆发目前已在全球感染了2000万人，其中美国感染了500万例（2020年8月11日）。迫切需要开发诊断和研究工具，以确定感染和疫苗效力。我们开发了多种血清学检测方法，使用新设计的SARS-CoV-2试剂检测血清中受体结合抗体的存在。第一种方法基于表面等离子体共振（SPR），可以在一次实验中定量测定抗体与SARS-CoV-2刺突蛋白的结合和对血管紧张素转换酶2 （ACE2）受体的阻断。第二种方法是基于酶联免疫吸附试验（ELISA）的方法，可以测量ACE2受体与SARS-CoV-2刺突蛋白的竞争和抗刺突抗体的阻断。该检测方法具有高度通用性，我们通过测量经实验性SARS-CoV-2疫苗免疫的小动物和非人灵长类动物血清的抗体功能，证明了该检测方法的广泛实用性。此外，我们采用该方法测量sars - cov -2感染患者血清的受体阻断。该试验显示与假病毒中和滴度相关。这种快速的替代中和诊断可广泛用于帮助研究SARS-CoV-2感染和评估疫苗的疗效。
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global pandemic of COVID-19, resulting in cases of mild to severe respiratory distress and significant mortality. The global outbreak of this novel coronavirus has now infected >20 million people worldwide, with >5 million cases in the United States (11 August 2020). The development of diagnostic and research tools to determine infection and vaccine efficacy is critically needed. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global pandemic of COVID-19, resulting in cases of mild to severe respiratory distress and significant mortality. The global outbreak of this novel coronavirus has now infected >20 million people worldwide, with >5 million cases in the United States (11 August 2020). The development of diagnostic and research tools to determine infection and vaccine efficacy is critically needed. We have developed multiple serologic assays using newly designed SARS-CoV-2 reagents for detecting the presence of receptor-binding antibodies in sera. The first assay is surface plasmon resonance (SPR) based and can quantitate both antibody binding to the SARS-CoV-2 spike protein and blocking to the Angiotensin-converting enzyme 2 (ACE2) receptor in a single experiment. The second assay is enzyme-linked immunosorbent assay (ELISA) based and can measure competition and blocking of the ACE2 receptor to the SARS-CoV-2 spike protein with antispike antibodies. The assay is highly versatile, and we demonstrate the broad utility of the assay by measuring antibody functionality of sera from small animals and nonhuman primates immunized with an experimental SARS-CoV-2 vaccine. In addition, we employ the assay to measure receptor blocking of sera from SARS-CoV-2-infected patients. The assay is shown to correlate with pseudovirus neutralization titers. This type of rapid, surrogate neutralization diagnostic can be employed widely to help study SARS-CoV-2 infection and assess the efficacy of vaccines.