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孤立 DNA 适体的合成及其在 AC 电热流基快速生物传感器中的应用,用于检测加标样品中的登革热病毒。

Synthesis of Isolated DNA Aptamer and Its Application of AC-Electrothermal Flow-Based Rapid Biosensor for the Detection of Dengue Virus in a Spiked Sample.

机构信息

Department of Chemical Engineering, Kwangwoon University, 20 Kwangwoon-Ro, Nowon-Gu, Seoul 01897, Republic of Korea.

Department of Electrical and Computer Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 08727, Republic of Korea.

出版信息

Bioconjug Chem. 2023 Aug 16;34(8):1486-1497. doi: 10.1021/acs.bioconjchem.3c00249. Epub 2023 Aug 1.

DOI:10.1021/acs.bioconjchem.3c00249
PMID:37527337
Abstract

Dengue fever is an infectious disease caused by the dengue virus (DENV) and is transmitted by mosquitoes in tropical and subtropical regions. The early detection method at a low cost is essential. To address this, we synthesized the isolated DENV aptamer for fabricating a rapid electrochemical biosensor on a Au interdigitated microgap electrode (AuIMGE). The DENV aptamers were generated using the SELEX (systemic evolution of ligands by exponential enrichment) method for binding to DENV surface envelope proteins. To reduce the manufacturing cost, unnecessary nucleotide sequences were excluded from the isolation process of the DENV aptamer. To reduce the detection time, the alternating current electrothermal flow (ACEF) technique was applied to the fabricated biosensor, which can shorten the detection time to 10 min. The performance of the biosensor was evaluated using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). In the diluted DENV protein solution, the linear range of the concentrations was from 1 pM to 1 μM and the LOD was 76.7 fM. Moreover, the proposed biosensor detected DENV in a diluted spiked sample at a linear range of 10 to 10 TCID/mL, while the detection performance was proven with an LOD of 1.74 × 10 TCIDmL along with high selectivity.

摘要

登革热是由登革病毒(DENV)引起的传染病,在热带和亚热带地区通过蚊子传播。因此,需要一种低成本的早期检测方法。为了解决这个问题,我们合成了分离的登革热病毒适配体,用于在 Au 叉指微间隙电极(AuIMGE)上制造快速电化学生物传感器。DENV 适配体是使用 SELEX(指数富集的配体系统进化)方法生成的,用于与 DENV 表面包膜蛋白结合。为了降低制造成本,从 DENV 适配体的分离过程中排除了不必要的核苷酸序列。为了缩短检测时间,将交流电热流(ACEF)技术应用于制造的生物传感器,这可以将检测时间缩短至 10 分钟。使用循环伏安法(CV)和电化学阻抗谱(EIS)评估了生物传感器的性能。在稀释的 DENV 蛋白溶液中,浓度的线性范围为 1 pM 至 1 μM,LOD 为 76.7 fM。此外,该生物传感器在稀释的加标样本中以 10 至 10 TCID/mL 的线性范围检测到 DENV,检测性能的 LOD 为 1.74×10 TCIDmL,具有高选择性。

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