Qiu Hongyu, Yuan Xin-Yong, Holloway Kimberly, Wood Heidi, Cabral Teresa, Grant Chris, McQueen Peter, Westmacott Garrett, Beniac Daniel R, Lin Lisa, Carpenter Michael, Kobasa Darwyn, Gräfenhan Tom, Cheney Ian Wayne
National Microbiology Laboratory, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB, R3E 3R2, Canada.
Heliyon. 2024 Jul 27;10(15):e35325. doi: 10.1016/j.heliyon.2024.e35325. eCollection 2024 Aug 15.
Rapid antigen test (RAT) is widely used for SARS-CoV-2 infection diagnostics. However, test sensitivity has decreased recently due to the emergence of the Omicron variant and its sublineages. Here we developed a panel of SARS-CoV-2 nucleocapsid protein (NP) specific mouse monoclonal antibodies (mAbs) and assessed their sensitivity and specificity to important SARS-CoV-2 variants. We identified seven mAbs that exhibited strong reactivity to SARS-CoV-2 variants and recombinant NP (rNP) by Western immunoblot or ELISA. Their specificity to SARS-CoV-2 was confirmed by negative or low reactivity to rNPs from SARS-CoV-1, MERS, and common human coronaviruses (HCoV-HKU1, HCoV-CO43, HCoV-NL63, and HCoV-229E). These seven mAbs were further tested by immunoplaque assay against selected variants of concern (VOCs), including two Omicron sublineages, and five mAbs (F461G13, F461G7, F459G7, F457G3, and F461G6), showed strong reactions, warranting further suitability testing for the development of diagnostic assay.
快速抗原检测（RAT）被广泛用于严重急性呼吸综合征冠状病毒2（SARS-CoV-2）感染的诊断。然而，由于奥密克戎变异株及其亚谱系的出现，最近检测灵敏度有所下降。在此，我们开发了一组针对SARS-CoV-2核衣壳蛋白（NP）的小鼠单克隆抗体（mAb），并评估了它们对重要SARS-CoV-2变异株的灵敏度和特异性。我们通过蛋白质免疫印迹法或酶联免疫吸附测定法鉴定出七种对SARS-CoV-2变异株和重组NP（rNP）表现出强反应性的mAb。它们对SARS-CoV-2的特异性通过对来自严重急性呼吸综合征冠状病毒1（SARS-CoV-1）、中东呼吸综合征冠状病毒（MERS）和常见人类冠状病毒（人冠状病毒HKU1、人冠状病毒OC43、人冠状病毒NL63和人冠状病毒229E）的rNP呈阴性或低反应性得以证实。通过免疫菌斑测定法对包括两个奥密克戎亚谱系在内的选定关注变异株（VOC）对这七种mAb进行了进一步测试，其中五种mAb（F461G13、F461G7、F459G7、F457G3和F461G6）表现出强烈反应，值得对其进行进一步的适用性测试以用于诊断检测的开发。
