用于 SARS-CoV-2 刺突糖蛋白检测的 DNA 适体选择。

DNA aptamer selection for SARS-CoV-2 spike glycoprotein detection.

机构信息

Laboratorio de Terapia Génica, Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del I.P.N., CDMX, 07360, Mexico.

Life and Medical Sciences (LIMES) Institute, University of Bonn, 53121, Bonn, Germany.

出版信息

Anal Biochem. 2022 May 15;645:114633. doi: 10.1016/j.ab.2022.114633. Epub 2022 Mar 2.

DOI:10.1016/j.ab.2022.114633

PMID:35247355

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8889740/

Abstract

The rapid spread of SARS-CoV-2 infection throughout the world led to a global public health and economic crisis triggering an urgent need for the development of low-cost vaccines, therapies and high-throughput detection assays. In this work, we used a combination of Ideal-Filter Capillary Electrophoresis SELEX (IFCE-SELEX), Next Generation Sequencing (NGS) and binding assays to isolate and validate single-stranded DNA aptamers that can specifically recognize the SARS-CoV-2 Spike glycoprotein. Two selected non-competing DNA aptamers, C7 and C9 were successfully used as sensitive and specific biological recognition elements for the development of electrochemical and fluorescent aptasensors for the SARS-CoV-2 Spike glycoprotein with detection limits of 0.07 fM and 41.87 nM, respectively.

摘要

SARS-CoV-2 感染在全球范围内迅速传播，导致全球公共卫生和经济危机，迫切需要开发低成本的疫苗、疗法和高通量检测方法。在这项工作中，我们使用理想过滤毛细管电泳 SELEX（IFCE-SELEX）、下一代测序（NGS）和结合测定法来分离和验证能够特异性识别 SARS-CoV-2 刺突糖蛋白的单链 DNA 适体。两个选定的非竞争 DNA 适体 C7 和 C9 成功地用作电化学和荧光适体传感器开发的敏感和特异性生物识别元件，用于检测 SARS-CoV-2 刺突糖蛋白，检测限分别为 0.07 fM 和 41.87 nM。