Development of a label-free impedimetric immunosensor for the detection of respiratory syncytial virus

IF 2.6 4区 化学 Q3 ELECTROCHEMISTRY
Tallita Stéfanne e Silva, Guilherme Ramos Oliveira e Freitas, Lucas Franco Ferreira, Diego Leoni Franco
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引用次数: 0

Abstract

Respiratory syncytial virus (RSV) is responsible for outbreaks of bronchiolitis, bronchitis, and pneumonia and is effective and lethal in children under five years old. Current RSV detection methods are expensive, time-consuming, and require highly skilled personnel. Therefore, we developed a cost-effective impedimetric immunosensor for the detection of RSV based on a pencil graphite electrode (PGE) modified with a polymeric film derived from 3-amino phenylacetic acid. The current responses in the potassium ferricyanide and methylene blue solutions suggest the presence of unchanged carboxylic acid groups in the polymer. With a similar number of electrons involved in the first step of the reaction and electron/proton ratio to those proposed for aniline, a mechanism for electropolymerization was proposed. The immunosensor was assembled by activating the carboxyl groups via an EDC/NHS nucleophilic substitution reaction with the amino groups present in the anti-RSV monoclonal antibody (mAb). The detection was performed using electrochemical impedance spectroscopy, and the data were evaluated using equivalent circuits. The charge transfer resistance values increased as PGE was modified in the following order: PGE < PGE/polymer < PGE/polymer/mAb < PGE/polymer/mAb/virus. 100 ng of mAb was used, and the immobilization times for mAb and blocker were 3 h and 50 min, respectively. The response time for RSV immobilization was 30 min. The limit of detection was 22.54 PFU mL−1, and the limit of quantification was 75.12 PFU mL−1, with a linear range from 50 – 2000 PFU mL−1, demonstrating that the proposed immunosensor is a viable alternative for the detection of RSV.

Graphical abstract

Abstract Image

Abstract Image

开发用于检测呼吸道合胞病毒的无标记阻抗免疫传感器
呼吸道合胞病毒(RSV)是导致支气管炎、气管炎和肺炎爆发的罪魁祸首,对五岁以下儿童有效且致命。目前的 RSV 检测方法成本高、耗时长,而且需要高技能人才。因此,我们开发了一种用于检测 RSV 的经济高效的阻抗免疫传感器,该传感器基于用 3-氨基苯乙酸衍生的聚合物薄膜修饰的铅笔石墨电极 (PGE)。铁氰化钾和亚甲基蓝溶液中的电流反应表明聚合物中存在未发生变化的羧酸基团。由于参与第一步反应的电子数和电子/质子比与苯胺的相似,因此提出了一种电聚合机理。通过 EDC/NHS 与抗 RSV 单克隆抗体(mAb)中的氨基发生亲核取代反应,激活羧基,从而组装出免疫传感器。使用电化学阻抗光谱进行检测,并使用等效电路对数据进行评估。电荷转移电阻值随着 PGE 的改性按以下顺序增加:PGE;PGE/聚合物;PGE/聚合物/mAb;PGE/聚合物/mAb/病毒。使用的 mAb 为 100 毫微克,mAb 和阻断剂的固定时间分别为 3 小时和 50 分钟。固定 RSV 的反应时间为 30 分钟。检测限为 22.54 PFU mL-1,定量限为 75.12 PFU mL-1,线性范围为 50 - 2000 PFU mL-1。
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来源期刊
CiteScore
4.80
自引率
4.00%
发文量
227
审稿时长
4.1 months
期刊介绍: The Journal of Solid State Electrochemistry is devoted to all aspects of solid-state chemistry and solid-state physics in electrochemistry. The Journal of Solid State Electrochemistry publishes papers on all aspects of electrochemistry of solid compounds, including experimental and theoretical, basic and applied work. It equally publishes papers on the thermodynamics and kinetics of electrochemical reactions if at least one actively participating phase is solid. Also of interest are articles on the transport of ions and electrons in solids whenever these processes are relevant to electrochemical reactions and on the use of solid-state electrochemical reactions in the analysis of solids and their surfaces. The journal covers solid-state electrochemistry and focusses on the following fields: mechanisms of solid-state electrochemical reactions, semiconductor electrochemistry, electrochemical batteries, accumulators and fuel cells, electrochemical mineral leaching, galvanic metal plating, electrochemical potential memory devices, solid-state electrochemical sensors, ion and electron transport in solid materials and polymers, electrocatalysis, photoelectrochemistry, corrosion of solid materials, solid-state electroanalysis, electrochemical machining of materials, electrochromism and electrochromic devices, new electrochemical solid-state synthesis. The Journal of Solid State Electrochemistry makes the professional in research and industry aware of this swift progress and its importance for future developments and success in the above-mentioned fields.
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