A Multiplex Microsphere IgG Assay for SARS-CoV-2 Using ACE2-Mediated Inhibition as a Surrogate for Neutralization.
A Multiplex Microsphere IgG Assay for SARS-CoV-2 Using ACE2-Mediated Inhibition as a Surrogate for Neutralization.
复制标题
使用ACE2介导的抑制作用作为中和替代的SARS-COV-2的多重微球IgG分析。
DOI：
10.1128/jcm.02489-20
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发表时间：
2021-01-21
影响因子：
9.4
通讯作者：
Pecora ND
中科院分区：
文献类型：
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作者：
Cameron A;Porterfield CA;Byron LD;Wang J;Pearson Z;Bohrhunter JL;Cardillo AB;Ryan-Muntz L;Sorensen RA;Caserta MT;Angeloni S;Hardy DJ;Zand MS;Pecora ND
2019冠状病毒病（COVID-19）大流行凸显了对新型病原体（如严重急性呼吸道综合征冠状病毒2（SARS-CoV-2））的血清学检测所固有的挑战。血清学检测可用于诊断和监测，但其有效性取决于其通量、灵敏度和特异性。在这里，我们描述了一种基于多重荧光微球的检测方法，3Flex，它可以检测三种主要SARS-CoV-2抗原-刺突蛋白（S），刺突ACE 2受体结合结构域（RBD）和核衣壳（NP）的抗体。2019冠状病毒病（COVID-19）大流行凸显了对新型病原体（如严重急性呼吸道综合征冠状病毒2（SARS-CoV-2））的血清学检测所固有的挑战。血清学检测可用于诊断和监测，但其有效性取决于其通量、灵敏度和特异性。在这里，我们描述了一种基于多重荧光微球的检测方法，3Flex，它可以检测三种主要SARS-CoV-2抗原-刺突蛋白（S），刺突ACE 2受体结合结构域（RBD）和核衣壳（NP）的抗体。使用213个大流行前样本评估特异性。使用来自125名独特患者的血清样本测量灵敏度，并与Abbott Architect SARS-CoV-2 IgG检测试剂盒进行比较，这些患者（n = 25）在5个时间间隔（症状发作后≤5、6 - 10、11 - 15、16 - 20和≥21天）中进行了相同分组。对于在症状发作≤5天获得的样本，3Flex检测试剂盒的灵敏度更高（48.0% vs 32.0%），但两种检测试剂盒使用症状发作≥21天获得的血清进行了重复检测。从通过图表审查表征COVID-19病程的患者（n = 140）中收集了大量（n = 534）丢弃的血清。这揭示了抗体应答的相对上升、峰值（S，23.8; RBD，23.6; NP，16.7 [症状发作后天数]）和下降。观察到相当大的人与人之间的变化与广泛抽样重症监护病房（ICU）患者的子集。使用可溶性ACE 2，证明了S和RBD的抗体结合抑制，而NP没有。综合这些数据，本研究描述了一种建立在灵活和高通量血清学平台上的检测方法的性能，该平台被证明适用于新感染因子的出现。
The coronavirus disease 2019 (COVID-19) pandemic has highlighted the challenges inherent to the serological detection of a novel pathogen such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Serological tests can be used diagnostically and for surveillance, but their usefulness depends on their throughput, sensitivity, and specificity. Here, we describe a multiplex fluorescent microsphere-based assay, 3Flex, that can detect antibodies to three major SARS-CoV-2 antigens—spike (S) protein, the spike ACE2 receptor-binding domain (RBD), and nucleocapsid (NP). The coronavirus disease 2019 (COVID-19) pandemic has highlighted the challenges inherent to the serological detection of a novel pathogen such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Serological tests can be used diagnostically and for surveillance, but their usefulness depends on their throughput, sensitivity, and specificity. Here, we describe a multiplex fluorescent microsphere-based assay, 3Flex, that can detect antibodies to three major SARS-CoV-2 antigens—spike (S) protein, the spike ACE2 receptor-binding domain (RBD), and nucleocapsid (NP). Specificity was assessed using 213 prepandemic samples. Sensitivity was measured and compared to that of the Abbott Architect SARS-CoV-2 IgG assay using serum samples from 125 unique patients equally binned (n = 25) into 5 time intervals (≤5, 6 to 10, 11 to 15, 16 to 20, and ≥21 days from symptom onset). With samples obtained at ≤5 days from symptom onset, the 3Flex assay was more sensitive (48.0% versus 32.0%), but the two assays performed comparably using serum obtained ≥21 days from symptom onset. A larger collection (n = 534) of discarded sera was profiled from patients (n = 140) whose COVID-19 course was characterized through chart review. This revealed the relative rise, peak (S, 23.8; RBD, 23.6; NP, 16.7 [in days from symptom onset]), and decline of the antibody response. Considerable interperson variation was observed with a subset of extensively sampled intensive care unit (ICU) patients. Using soluble ACE2, inhibition of antibody binding was demonstrated for S and RBD, and not for NP. Taking the data together, this study described the performance of an assay built on a flexible and high-throughput serological platform that proved adaptable to the emergence of a novel infectious agent.