Luminex Corporation.
Departments of Pathology and Laboratory Medicine, Clinical Microbiology, University of Rochester Medical Center.
J Vis Exp. 2021 Apr 6(170). doi: 10.3791/62487.
The COVID-19 pandemic has underscored the need for rapid high-throughput methods for sensitive and specific serological detection of infection with novel pathogens, such as SARS-CoV-2. Multiplex serological testing can be particularly useful because it can simultaneously analyze antibodies to multiple antigens that optimizes pathogen coverage, and controls for variability in the organism and the individual host response. Here we describe a SARS-CoV-2 IgG 3-plex fluorescent microsphere-based assay that can detect both IgM and IgG antibodies to three major SARS-CoV-2 antigens-the spike (S) protein, spike angiotensin-converting enzyme-2 (ACE2) receptor-binding domain (RBD), and nucleocapsid (Nc). The assay was shown to have comparable performance to a SARS-CoV-2 reference assay for IgG in serum obtained at ≥21 days from symptom onset but had higher sensitivity with samples collected at ≤5 days from symptom onset. Further, using soluble ACE2 in a neutralization assay format, inhibition of antibody binding was demonstrated for S and RBD.
COVID-19 大流行凸显了对新型病原体（如 SARS-CoV-2）进行敏感和特异性血清学检测的快速高通量方法的需求。多重血清学检测特别有用，因为它可以同时分析针对多种抗原的抗体，从而优化病原体的覆盖范围，并控制生物体和个体宿主反应的变异性。在这里，我们描述了一种基于 SARS-CoV-2 IgG 三plex 荧光微球的检测方法，该方法可以检测到三种主要的 SARS-CoV-2 抗原的 IgM 和 IgG 抗体 - 刺突（S）蛋白、刺突血管紧张素转换酶 2（ACE2）受体结合域（RBD）和核衣壳（Nc）。该检测方法在≥21 天从症状发作时获得的血清中与 SARS-CoV-2 参考检测方法相比具有相当的 IgG 性能，但在≤5 天从症状发作时采集的样本中具有更高的敏感性。此外，使用可溶性 ACE2 在中和测定格式中，证明了 S 和 RBD 抗体结合的抑制。
