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无标记电化学适体传感器的研制及其用于裂谷热病毒的检测。

Development of a label-free electrochemical aptasensor for Rift Valley fever virus detection.

机构信息

Department of Chemistry, Alfaisal University, Al Zahrawi Street, Al Maather, Al Takhassusi Road, Riyadh, 11533, Saudi Arabia.

Bioengineering Laboratory, Higher National School of Biotechnology, Constantine, Algeria.

出版信息

Sci Rep. 2024 Oct 12;14(1):23892. doi: 10.1038/s41598-024-74314-7.

Abstract

In this research, we describe the first aptasensor for the detection of the Rift Valley Fever virus (RVFV). The process involved the selection of aptamers through the systematic evolution of ligands by the exponential enrichment (SELEX) technique. After 12 rounds of selection, 6 aptamers were selected and the corresponding binding affinities were assessed using fluorescence binding assays, revealing dissociation constants ranging from 15.45 to 40.98 nM. Notably, among the aptamers, RV2 and RV3 exhibited the highest binding affinities toward RVFV, with dissociation constants of 15.45 and 18.62 nM, respectively. Thiol-modified aptamers were subsequently immobilized onto screen-printed gold electrodes, facilitating the label-free detection of RVFV through square wave voltammetry. The voltammetric aptasensor demonstrated an excellent sensitivity, with a detection limit of 0.015 ng/mL. In addition, cross-reactivity assessments were conducted, where negligible response was obtained when the aptasensor was exposed to non-specific proteins.

摘要

在这项研究中,我们描述了第一个用于检测裂谷热病毒(RVFV)的适体传感器。该过程涉及通过指数富集(SELEX)技术选择配体的适体。经过 12 轮选择,选出了 6 个适体,并使用荧光结合分析评估了它们的相应结合亲和力,发现解离常数范围在 15.45 到 40.98 nM 之间。值得注意的是,在这些适体中,RV2 和 RV3 对 RVFV 的结合亲和力最高,解离常数分别为 15.45 和 18.62 nM。随后,巯基修饰的适体被固定在丝网印刷金电极上,通过方波伏安法实现了对 RVFV 的无标记检测。该伏安适体传感器表现出优异的灵敏度,检测限为 0.015ng/mL。此外,还进行了交叉反应性评估,当适体传感器暴露于非特异性蛋白质时,几乎没有得到响应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2e3/11470950/8a577a4d58bb/41598_2024_74314_Fig1_HTML.jpg

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