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基于电场辅助的均相杂交磁生物传感器用于核酸的超快检测。

A homogeneous hybridization magnetic biosensor based on electric field assistance for ultrafast nucleic acid detection.

机构信息

Shandong Provincial Key Laboratory of Biochemical Engineering, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, College of Marine Science and Biological Engineering, Qingdao University of Science and Technology, Qingdao, 266042, PR China.

College of Chemistry and Molecular Engineering, State Key Laboratory Base of Eco-Chemical Engineering, International Science and Technology Cooperation Base of Eco-chemical Engineering and Green Manufacturing, Qingdao University of Science and Technology, Qingdao 266042, China.

出版信息

Anal Methods. 2023 Jul 13;15(27):3325-3332. doi: 10.1039/d3ay00548h.

Abstract

Electrochemical biosensing is a sensitive strategy widely used in the field of nucleic acid detection. However, electrochemical biosensors generally involve time-consuming and labor-intensive probe immobilization processes. In this study, an electrochemical DNA biosensor based on homogeneous hybridization in solution was designed for nucleic acid detection without probe immobilization, which is different from most biosensors. The capture probe, detection probe, and target DNA were hybridized rapidly under an electric field to form a "sandwich" structure within 90 s, and the "sandwich" hybrid could be specifically coupled to streptavidin-modified magnetic beads within 5 min. Finally, the magnetic beads were enriched by using polypyrrole (PPy)/carbon nanotube (CNT)-modified magnetic electrodes and the signal was detected by differential pulse voltammetry (DPV). The magnetic biosensor constructed in this study could detect targets over a good linear dynamic range spanning 100 pM to 100 nM in 400 s, while those involving conventional hybridization methods always take 2 h or more. Because of the specific binding of streptavidin and biotin, this strategy showed high specificity. Taken together, the homogenous hybridization magnetic biosensor constructed with electric field assistance presents a potential diagnostic method for rapid DNA detection and provides a new idea for rapid nucleic acid detection in clinical practice.

摘要

电化学生物传感是一种广泛应用于核酸检测领域的灵敏策略。然而,电化学生物传感器通常涉及耗时耗力的探针固定化过程。在本研究中,设计了一种基于溶液中均相杂交的电化学 DNA 生物传感器,用于无需探针固定化的核酸检测,这与大多数生物传感器不同。在电场作用下,捕获探针、检测探针和目标 DNA 迅速杂交,在 90 秒内形成“三明治”结构,“三明治”杂交体在 5 分钟内可特异性结合链霉亲和素修饰的磁珠。最后,采用聚吡咯(PPy)/碳纳米管(CNT)修饰的磁性电极富集磁珠,并通过差分脉冲伏安法(DPV)检测信号。本研究构建的磁性生物传感器可在 400 秒内检测到 100 pM 至 100 nM 的良好线性动态范围内的靶标,而涉及常规杂交方法的传感器通常需要 2 小时或更长时间。由于链霉亲和素和生物素的特异性结合,该策略表现出高特异性。总之,电场辅助的均相杂交磁性生物传感器为快速 DNA 检测提供了一种潜在的诊断方法,并为临床实践中的快速核酸检测提供了新的思路。

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