Oswaldo Cruz Foundation (FIOCRUZ), Center for Technological Development in Health (CDTS)/National Institute of Science and Technology for Innovation in Neglected Populations Diseases (INCT-IDPN), Rio de Janeiro 21040-900, RJ, Brazil.
Laboratory of Epidemiology and Molecualr Systematics (LESM), Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro 21040-900, RJ, Brazil.
Biosensors (Basel). 2022 Apr 26;12(5):272. doi: 10.3390/bios12050272.
The coronavirus disease of 2019 (COVID-19) is caused by an infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). It was recognized in late 2019 and has since spread worldwide, leading to a pandemic with unprecedented health and financial consequences. There remains an enormous demand for new diagnostic methods that can deliver fast, low-cost, and easy-to-use confirmation of a SARS-CoV-2 infection. We have developed an affordable electrochemical biosensor for the rapid detection of serological immunoglobulin G (IgG) antibody in sera against the spike protein.
A previously identified linear B-cell epitope (EP) specific to the SARS-CoV-2 spike glycoprotein and recognized by IgG in patient sera was selected for the target molecule. After synthesis, the EP was immobilized onto the surface of the working electrode of a commercially available screen-printed electrode (SPE). The capture of SARS-CoV-2-specific IgGs allowed the formation of an immunocomplex that was measured by square-wave voltammetry from its generation of hydroquinone (HQ).
An evaluation of the performance of the EP-based biosensor presented a selectivity and specificity for COVID-19 of 93% and 100%, respectively. No cross-reaction was observed to antibodies against other diseases that included Chagas disease, Chikungunya, Leishmaniosis, and Dengue. Differentiation of infected and non-infected individuals was possible even at a high dilution factor that decreased the required sample volumes to a few microliters.
The final device proved suitable for diagnosing COVID-19 by assaying actual serum samples, and the results displayed good agreement with the molecular biology diagnoses. The flexibility to conjugate other EPs to SPEs suggests that this technology could be rapidly adapted to diagnose new variants of SARS-CoV-2 or other pathogens.
2019 年冠状病毒病(COVID-19)是由严重急性呼吸系统综合征冠状病毒 2(SARS-CoV-2)感染引起的。它于 2019 年末被发现,此后已在全球范围内传播,导致了前所未有的健康和经济后果的大流行。仍然需要新的诊断方法来快速、低成本和方便地确认 SARS-CoV-2 感染。我们开发了一种经济实惠的电化学生物传感器,用于快速检测血清中针对刺突蛋白的血清学免疫球蛋白 G(IgG)抗体。
选择针对 SARS-CoV-2 刺突糖蛋白的先前鉴定的线性 B 细胞表位(EP),并针对患者血清中的 IgG 进行识别。合成后,EP 被固定在市售的丝网印刷电极(SPE)的工作电极表面上。SARS-CoV-2 特异性 IgGs 的捕获允许形成免疫复合物,通过方波伏安法从其产生的对苯二酚(HQ)来测量。
基于 EP 的生物传感器的性能评估显示,COVID-19 的选择性和特异性分别为 93%和 100%。对包括恰加斯病、基孔肯雅热、利什曼病和登革热在内的其他疾病的抗体没有交叉反应。即使在高稀释因子下也可以区分感染和未感染的个体,该稀释因子降低了所需的样本量至几微升。
最终设备通过检测实际的血清样本证明适合诊断 COVID-19,并且结果与分子生物学诊断显示出良好的一致性。将其他 EP 共轭到 SPE 上的灵活性表明,该技术可以快速适应诊断 SARS-CoV-2 或其他病原体的新变体。