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用于即时检测 SARS-CoV-2(COVID-19)的超快速可回收 DNA 生物传感器。

Ultra-fast and recyclable DNA biosensor for point-of-care detection of SARS-CoV-2 (COVID-19).

机构信息

College of Pharmacy, Ajou University, Suwon 16499, South Korea.

Electronic Engineering, Kwangwoon University, Seoul 01897, South Korea.

出版信息

Biosens Bioelectron. 2021 Aug 1;185:113177. doi: 10.1016/j.bios.2021.113177. Epub 2021 Mar 24.

Abstract

Rapid diagnosis and case isolation are pivotal to controlling the current pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, a label-free DNA capacitive biosensor for the detection of SARS-CoV-2 that demonstrates real-time, low-cost, and high-throughput screening of nucleic acid samples is presented. Our novel biosensor composed of the interdigitated platinum/titanium electrodes on the glass substrate can detect the hybridization of analyte DNA with probe DNA. The hybridization signals of specific DNA sequences were verified through exhaustive physicochemical analytical techniques such as Fourier transform infrared (FT-IR) spectrometry, contact-angle analysis, and capacitance-frequency measurements. For a single-step hybridized reaction, the fabricated kit exhibited significant sensitivity (capacitance change, ΔC = ~2 nF) in detecting the conserved region of the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) gene with high sensitivity of 0.843 nF/nM. In addition to capacitive measurements, this selective detection was confirmed by the fluorescence image and intensity from a SARS-CoV-2 gene labeled with a fluorescent dye. We also demonstrated that the kits are recyclable by surface ozone treatment using UV irradiation. Thus, these kits could potentially be applied to various types of label-free DNA, thereby acting as rapid, cost-effective biosensors for several diseases.

摘要

快速诊断和病例隔离是控制当前严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 大流行的关键。在这项研究中,我们提出了一种用于检测 SARS-CoV-2 的无标记 DNA 电容式生物传感器,该传感器可实时、低成本、高通量筛选核酸样本。我们的新型生物传感器由玻璃衬底上的叉指式铂/钛电极组成,可以检测分析物 DNA 与探针 DNA 的杂交。通过傅里叶变换红外 (FT-IR) 光谱、接触角分析和电容-频率测量等详尽的物理化学分析技术,验证了特定 DNA 序列的杂交信号。对于单步杂交反应,所制备的试剂盒在检测 SARS-CoV-2 RNA 依赖性 RNA 聚合酶 (RdRp) 基因保守区时表现出显著的灵敏度(电容变化,ΔC~2 nF),灵敏度高达 0.843 nF/nM。除了电容测量外,通过用荧光染料标记的 SARS-CoV-2 基因的荧光图像和强度也证实了这种选择性检测。我们还证明,试剂盒可以通过使用紫外线辐照的表面臭氧处理进行回收利用。因此,这些试剂盒有可能应用于各种类型的无标记 DNA,从而成为用于多种疾病的快速、经济有效的生物传感器。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03b9/7987504/40a9c64a315a/gr1_lrg.jpg

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