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一次性氧化石墨烯丝网印刷电极与便携式设备集成,用于检测临床样本中的 SARS-CoV-2。

Disposable graphene-oxide screen-printed electrode integrated with portable device for detection of SARS-CoV-2 in clinical samples.

机构信息

DBT-National Institute of Animal Biotechnology (NIAB), Hyderabad 500032, Telangana, India; DBT-Regional Centre for Biotechnology (RCB), Faridabad 121001, Haryana, India.

DBT-National Institute of Animal Biotechnology (NIAB), Hyderabad 500032, Telangana, India.

出版信息

Bioelectrochemistry. 2024 Aug;158:108722. doi: 10.1016/j.bioelechem.2024.108722. Epub 2024 Apr 27.

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnosis is the need of the hour, as cases are persistently increasing, and new variants are constantly emerging. The ever-changing nature of the virus leading to multiple variants, has brought an imminent need for early, accurate and rapid detection methods. Herein, we have reported the design and fabrication of Screen-Printed Electrodes (SPEs) with graphene oxide (GO) as working electrode and modified with specific antibodies for SARS-CoV-2 Receptor Binding Domain (RBD). Flexibility of design, and portable nature has made SPEs the superior choice for electrochemical analysis. The developed immunosensor can detect RBD as low as 0.83 fM with long-term storage capacity. The fabricated SPEs immunosensor was tested using a miniaturized portable device and potentiostat on 100 patient nasopharyngeal samples and corroborated with RT-PCR data, displayed 94 % sensitivity. Additionally, the in-house developed polyclonal antibodies detected RBD antigen of the mutated Omicron variant of SARS-CoV-2 successfully. We have not observed any cross-reactivity/binding of the fabricated immunosensor with MERS (cross-reactive antigen) and Influenza A H1N1 (antigen sharing common symptoms). Hence, the developed SPEs sensor may be applied for bedside point-of-care diagnosis of SARS-CoV-2 using miniaturized portable device, in clinical samples.

摘要

严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 的诊断是当务之急,因为病例持续增加,新的变体不断出现。病毒不断变化的性质导致了多种变体的出现,这就迫切需要早期、准确和快速的检测方法。在这里,我们报告了丝网印刷电极 (SPEs) 的设计和制造,其工作电极采用氧化石墨烯 (GO) ,并用针对 SARS-CoV-2 受体结合域 (RBD) 的特异性抗体进行修饰。设计的灵活性和便携性使 SPEs 成为电化学分析的首选。所开发的免疫传感器可以检测到低至 0.83 fM 的 RBD,并且具有长期存储能力。使用小型化便携式设备和电化学工作站对 100 个鼻咽患者样本进行了测试,并与 RT-PCR 数据进行了比较,显示出 94%的灵敏度。此外,内部开发的多克隆抗体成功检测到 SARS-CoV-2 奥密克戎变体的 RBD 抗原。我们没有观察到所制造的免疫传感器与中东呼吸综合征 (MERS) (交叉反应性抗原) 和甲型 H1N1 流感 (具有共同症状的抗原) 之间有任何交叉反应/结合。因此,所开发的 SPEs 传感器可以用于使用小型化便携式设备在临床样本中进行床边即时 SARS-CoV-2 诊断。

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