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低成本棉拭子电化学免疫传感器的研制及其用于 SARS-CoV-2 的检测。

Development of a Low-Cost Cotton-Tipped Electrochemical Immunosensor for the Detection of SARS-CoV-2.

机构信息

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

King Faisal Specialist Hospital and Research Centre, Zahrawi Street, Al Maather, Riyadh 12713, Saudi Arabia.

出版信息

Anal Chem. 2021 Jan 26;93(3):1826-1833. doi: 10.1021/acs.analchem.0c04719. Epub 2020 Dec 28.

Abstract

Collection of nasopharyngeal samples using swabs followed by the transfer of the virus into a solution and an RNA extraction step to perform reverse transcription polymerase chain reaction (PCR) is the primary method currently used for the diagnosis of COVID-19. However, the need for several reagents and steps and the high cost of PCR hinder its worldwide implementation to contain the outbreak. Here, we report a cotton-tipped electrochemical immunosensor for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus antigen. Unlike the reported approaches, we integrated the sample collection and detection tools into a single platform by coating screen-printed electrodes with absorbing cotton padding. The immunosensor was fabricated by immobilizing the virus nucleocapsid (N) protein on carbon nanofiber-modified screen-printed electrodes which were functionalized by diazonium electrografting. The detection of the virus antigen was achieved via swabbing followed by competitive assay using a fixed amount of N protein antibody in the solution. A square wave voltammetric technique was used for the detection. The limit of detection for our electrochemical biosensor was 0.8 pg/mL for SARS-CoV-2, indicating very good sensitivity for the sensor. The biosensor did not show significant cross-reactivity with other virus antigens such as influenza A and HCoV, indicating high selectivity of the method. Moreover, the biosensor was successfully applied for the detection of the virus antigen in spiked nasal samples showing excellent recovery percentages. Thus, our electrochemical immunosensor is a promising diagnostic tool for the direct rapid detection of the COVID-19 virus that requires no sample transfer or pretreatment.

摘要

采集鼻咽样本,使用拭子将病毒转移到溶液中,然后进行 RNA 提取步骤,以进行逆转录聚合酶链反应(PCR),这是目前用于诊断 COVID-19 的主要方法。然而,需要多种试剂和步骤,以及 PCR 的高成本,阻碍了其在全球范围内的实施,以遏制疫情的爆发。在这里,我们报告了一种用于检测严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)病毒抗原的棉签电化学免疫传感器。与已报道的方法不同,我们通过将吸收性棉签垫涂覆在丝网印刷电极上,将样本采集和检测工具集成到单个平台中。通过将病毒核衣壳(N)蛋白固定在经过叠氮电接枝功能化的碳纤维纳米纤维修饰的丝网印刷电极上,制备了免疫传感器。通过拭子擦拭后,使用溶液中固定量的 N 蛋白抗体进行竞争测定,从而实现了对病毒抗原的检测。采用方波伏安技术进行检测。我们的电化学生物传感器对 SARS-CoV-2 的检测限为 0.8pg/mL,表明传感器具有非常好的灵敏度。该生物传感器与其他病毒抗原(如流感 A 和 HCoV)没有明显的交叉反应性,表明该方法具有很高的选择性。此外,该生物传感器成功地应用于检测加标鼻样本中的病毒抗原,显示出极好的回收率。因此,我们的电化学免疫传感器是一种有前途的诊断工具,可用于直接快速检测 COVID-19 病毒,无需样品转移或预处理。

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