Minagawa Hirotaka, Sawa Hirofumi, Fujita Tomoko, Kato Shintaro, Inaguma Asumi, Hirose Miwako, Orba Yasuko, Sasaki Michihito, Tabata Koshiro, Nomura Naoki, Shingai Masashi, Suzuki Yasuhiko, Horii Katsunori
NEC Solution Innovators, Ltd., 1-18-7, Shinkiba, Koto-ku, Tokyo, 136-8627, Japan.
Division of Molecular Pathobiology, International Institute for Zoonosis Control, Hokkaido University, N20, W10, Kita-ku, Sapporo, 001-0020, Japan; International Collaboration Unit, International Institute for Zoonosis Control, Hokkaido University, N20, W10, Kita-ku, Sapporo, 001-0020, Japan; One Health Research Center, Hokkaido University, N20, W10, Kita-ku, Sapporo, 001-0020, Japan.
Biochem Biophys Res Commun. 2022 Jul 23;614:207-212. doi: 10.1016/j.bbrc.2022.04.071. Epub 2022 May 2.
Simple, highly sensitive detection technologies for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are crucial for the effective implementation of public health policies. We used the systematic evolution of ligands by exponential enrichment with a modified DNA library, including a base-appended base (uracil with a guanine base at its fifth position), to create an aptamer with a high affinity for the receptor-binding domain (RBD) of the SARS-CoV-2 spike glycoprotein. The aptamer had a dissociation constant of 1.2 and < 1 nM for the RBD and spike trimer, respectively. Furthermore, enzyme-linked aptamer assays confirmed that the aptamer binds to isolated authentic SARS-CoV-2 wild-type and B.1.617.2 (delta variant). The binding signal was larger that of commercially available anti-SARS-CoV-2 RBD antibody. Thus, this aptamer as a sensing element will enable the highly sensitive detection of SARS-CoV-2.
用于严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的简单、高灵敏度检测技术对于有效实施公共卫生政策至关重要。我们使用经过修饰的DNA文库（包括碱基附加碱基，即第五位带有鸟嘌呤碱基的尿嘧啶）通过指数富集配体系统进化技术，来创建对SARS-CoV-2刺突糖蛋白的受体结合域（RBD）具有高亲和力的适体。该适体对RBD和刺突三聚体的解离常数分别为1.2和<1 nM。此外，酶联适体分析证实该适体可与分离出的真实SARS-CoV-2野生型和B.1.617.2（德尔塔变体）结合。其结合信号比市售抗SARS-CoV-2 RBD抗体的信号更强。因此，这种作为传感元件的适体将能够实现对SARS-CoV-2的高灵敏度检测。
