Department of Immunology, Federal University of Triângulo Mineiro, Uberaba 38025-180, Brazil.
INCT-Neuroimmune Modulation, Uberaba 38025-350, Brazil.
Biosensors (Basel). 2024 Oct 22;14(11):514. doi: 10.3390/bios14110514.
The COVID-19 pandemic, caused by the SARS-CoV-2 virus, has posed a major challenge to global health. The development of fast, accurate, and accessible diagnostic methods is essential in controlling the disease and mitigating its impacts. In this context, electrochemical biosensors present themselves as promising tools for the efficient monitoring of SARS-CoV-2 infection. We have developed a highly specific biosensor for the detection of anti-SARS-CoV-2 antibodies in patient sera. The use of the RBD-S region as an antigen, although purified to minimize cross-linking, poses a specific challenge. The structural similarity between SARS-CoV-2 and other respiratory viruses, as well as the complexity of the serum matrix, hinders robust analytical strategies to ensure diagnostic accuracy. This work presents a novel immunosensor for COVID-19 diagnosis using laser-induced graphene (LIG) electrodes subjected to electrochemical reduction with graphene (named rGraphene-LIG). In the present study, we chose an initial approach focused on demonstrating the concept and evaluating the feasibility of the rGraphene-LIG sensor for SARS-CoV-2 detection. The rGraphene-LIG electrodes presented a notable current increase for the redox probe in the aqueous solution of a mixture of 5 mmol L potassium ferricyanide/ferrocyanide ([Fe(CN)]/[Fe(CN)]) in 0.1 mol L KCl (pH set at 7.4). As a proof of concept, the rGraphene-LIG electrode was applied for antibody determination in real samples using cyclic voltammetry, and a limit of detection (LOD) of 0.032 μg L was achieved. When determining antigens in commercial samples, we obtained an LOD of 560 ηg mL and a limit of quantification of 1677 ηg mL. The results of the electrochemical experiments were in accordance with the surface roughness obtained from atomic force microscopy images. Based on these results, the rGraphene-LIG electrode is shown to be an excellent platform for immunoglobulin detection when present in individuals after antigenic exposure caused by SARS-CoV-2.
新型冠状病毒肺炎(COVID-19)由 SARS-CoV-2 病毒引起,对全球健康构成重大挑战。开发快速、准确和易于使用的诊断方法对于控制疾病和减轻其影响至关重要。在这种情况下,电化学生物传感器作为监测 SARS-CoV-2 感染的有效工具具有广阔的应用前景。我们开发了一种用于检测患者血清中抗 SARS-CoV-2 抗体的高特异性生物传感器。虽然 RBD-S 区域作为抗原被纯化以尽量减少交联,但仍存在特定的挑战。SARS-CoV-2 与其他呼吸道病毒之间的结构相似性以及血清基质的复杂性,阻碍了开发稳健的分析策略来确保诊断的准确性。本工作提出了一种使用激光诱导石墨烯(LIG)电极的新型 COVID-19 诊断免疫传感器,该电极经过电化学还原处理并用石墨烯(命名为 rGraphene-LIG)修饰。在本研究中,我们选择了一种初始方法,专注于证明 rGraphene-LIG 传感器用于 SARS-CoV-2 检测的概念和可行性。rGraphene-LIG 电极在含有 5 mmol L 铁氰化钾/亚铁氰化钾([Fe(CN)]/[Fe(CN)])的水溶液和 0.1 mol L KCl(pH 值设定为 7.4)的混合溶液中的氧化还原探针的电流显著增加。作为概念验证,使用循环伏安法将 rGraphene-LIG 电极应用于实际样品中的抗体测定,检测限(LOD)达到 0.032 μg L。在确定商业样品中的抗原时,我们获得了 560 ηg mL 的 LOD 和 1677 ηg mL 的定量限。电化学实验的结果与原子力显微镜图像得到的表面粗糙度相符。基于这些结果,rGraphene-LIG 电极在 SARS-CoV-2 抗原暴露后个体中存在时,被证明是一种用于免疫球蛋白检测的优秀平台。