一种具有碱基附加修饰DNA的高亲和力适配体与分离出的野生型和B.1.617.2（德尔塔变体）真实新冠病毒毒株结合。
A high-affinity aptamer with base-appended base-modified DNA bound to isolated authentic SARS-CoV-2 strains wild-type and B.1.617.2 (delta variant).
作者信息
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
机构信息
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.
出版信息
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。
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