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临床样本中新型冠状病毒2019(SARS-CoV-2)的超灵敏伏安检测

Ultrasensitive voltammetric detection of SARS-CoV-2 in clinical samples.

作者信息

Kowalczyk Agata, Kasprzak Artur, Ruzycka-Ayoush Monika, Podsiadły Edyta, Demkow Urszula, Grudzinski Ireneusz P, Nowicka Anna M

机构信息

Faculty of Chemistry, University of Warsaw, Pasteura 1 Str., PL 02-093 Warsaw, Poland.

Faculty of Chemistry, Warsaw University of Technology, Nowakowskiego 3 Str., PL 00-664 Warsaw, Poland.

出版信息

Sens Actuators B Chem. 2022 Nov 15;371:132539. doi: 10.1016/j.snb.2022.132539. Epub 2022 Aug 23.

Abstract

In every pandemic, it is critical to test as many people as possible and keep track of the number of new cases of infection. Therefore, there is a need for novel, fast and unambiguous testing methods. In this study, we designed a sandwich-type voltammetric immunosensor based on unlabeled- and labeled with a redox probe antibodies against virus spike protein for fast and ultrasensitive detection of SARS-CoV-2. The process of the preparation of the sensor layer included chemisorption of cysteamine layer and covalent anchoring of antibody specific for the S1 subunit of the S protein. The source of the voltametric signal was the antibody labeled with the redox probe, which was introduced onto biosensor surface only after the recognition of the virus. This easy-to-handle immunosensor was characterized by a wide analytical range (2.0·10 to 0.20 mg·L) and low detection limit (8.0·10 mg·L ≡ 0.08 pg·mL ≡ 4 virions·μL). The utility of the designed device was also evidenced by the detection of SARS-CoV-2 in the clinical samples. Moreover, the main advantage and a huge novelty of the developed device, compared to those already existing, is the moment of generating the analytical signal of the redox probe that appears only after the virus recognition. Thus, our diagnostic innovation may considerably contribute to controlling the COVID-19 pandemic. The as-developed immunosensor may well offer a novel alternative approach for viral detection that could complement or even replace the existing methods.

摘要

在每一次大流行中,尽可能多地检测人群并跟踪新感染病例数量至关重要。因此,需要新颖、快速且明确的检测方法。在本研究中,我们设计了一种基于未标记和用氧化还原探针标记的针对病毒刺突蛋白抗体的夹心型伏安免疫传感器,用于快速、超灵敏地检测严重急性呼吸综合征冠状病毒2(SARS-CoV-2)。传感器层的制备过程包括半胱胺层的化学吸附和对S蛋白S1亚基具有特异性的抗体的共价固定。伏安信号的来源是用氧化还原探针标记的抗体,该抗体仅在识别病毒后才被引入生物传感器表面。这种易于操作的免疫传感器具有宽分析范围(2.0·10至0.20 mg·L)和低检测限(8.0·10 mg·L≡0.08 pg·mL≡4个病毒粒子·μL)的特点。所设计装置的实用性还通过在临床样本中检测SARS-CoV-2得到了证明。此外,与现有装置相比,所开发装置的主要优点和巨大新颖之处在于氧化还原探针的分析信号仅在病毒识别后才产生。因此,我们的诊断创新可能会对控制2019冠状病毒病大流行做出重大贡献。所开发的免疫传感器很可能为病毒检测提供一种新颖的替代方法,可补充甚至取代现有方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc39/9395233/b023e2778739/gr2_lrg.jpg

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