Oak Ridge Institute for Science and Education, Oak Ridge Associated Universities, 100 ORAU way, Oak Ridge, TN, 37830, USA.
National Center for Immunization and Respiratory Diseases, Division of Viral Diseases, Gastroenteritis and Respiratory Viruses Laboratory Branch, Centers for Disease Control and Prevention 1600 Clifton Rd NE, Atlanta, GA, 30329, USA.
Sci Rep. 2019 Feb 4;9(1):1390. doi: 10.1038/s41598-018-37747-5.
Known human coronaviruses (hCoV) usually cause mild to moderate upper-respiratory tract illnesses, except SARS-CoV and MERS-CoV, which, in addition to mild illness can also be associated with severe respiratory diseases and high mortality rates. Well-characterized multiplexed serologic assays are needed to aid in rapid detection and surveillance of hCoVs. The present study describes development and evaluation of a multiplexed magnetic microsphere immunoassay (MMIA) to simultaneously detect immunoglobulin G (IgG) antibodies specific for recombinant nucleocapsid proteins (recN) from hCoVs 229E, NL63, OC43, HKU1, SARS-CoV, and MERS-CoV. We used paired human sera to screen for IgG with reactivity against six hCoVs to determine assay sensitivity, specificity and reproducibility. We found no signal interference between monoplex and multiplex assay formats (R range = 0.87-0.97). Screening of paired human sera using MMIA, resulted in 92 of 106 (sensitivity: 86%) as positive and 68 of 80 (specificity: 84%) as negative. This study serves as a proof of concept that it is feasible to develop and use a multiplexed microsphere immunoassay as a next generation screening tool for use in large scale seroprevalence studies of hCoVs.
已知的人类冠状病毒（hCoV）通常会引起轻度至中度上呼吸道疾病，除了 SARS-CoV 和 MERS-CoV，这两种病毒除了轻度疾病外，还可能与严重的呼吸道疾病和高死亡率有关。需要经过充分验证的多重血清学检测方法来帮助快速检测和监测 hCoVs。本研究描述了一种多重磁珠免疫分析（MMIA）的开发和评估，用于同时检测针对 hCoVs 229E、NL63、OC43、HKU1、SARS-CoV 和 MERS-CoV 的重组核衣壳蛋白（recN）的 IgG 抗体。我们使用配对的人血清来筛选针对六种 hCoVs 的 IgG，以确定检测的灵敏度、特异性和重现性。我们发现单重和多重检测方法之间没有信号干扰（R 范围=0.87-0.97）。使用 MMIA 对配对的人血清进行筛选，结果 106 份血清中有 92 份（灵敏度：86%）为阳性，80 份中有 68 份（特异性：84%）为阴性。本研究证明了开发和使用多重微球免疫分析作为下一代筛查工具用于大规模 hCoV 血清流行率研究是可行的。
