Max Planck Tandem Group in Nanobioengineering, Institute of Chemistry, Faculty of Natural and Exact Sciencies, University of Antioquia, Complejo Ruta N, Calle 67 N° 52-20, Medellín, 050010, Colombia.
Programa de Estudio y Control de Enfermedades Tropicales (PECET), Facultad de Medicina, Universidad de Antioquia, Calle 62 N° 52-59, Medellín, Colombia.
Talanta. 2022 Aug 1;245:123482. doi: 10.1016/j.talanta.2022.123482. Epub 2022 Apr 16.
Infection caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is responsible for the Coronavirus disease (COVID-19) and the current pandemic. Its mortality rate increases, demonstrating the imperative need for acute and rapid diagnostic tools as an alternative to current serological tests and molecular techniques. Features of electrochemical genosensor devices make them amenable for fast and accurate testing closer to the patient. This work reports on a specific electrochemical genosensor for SARS-CoV-2 detection and discrimination against homologous respiratory viruses. The electrochemical biosensor was assembled by immobilizing thiolated capture probes on top of maleimide-coated magnetic particles, followed by specific target hybridization between the capture and biotinylated signaling probes in a sandwich-type manner. The probes were rigorously designed bioinformatically and tested in vitro. Enzymatic complexes based on streptavidin-horseradish peroxidase linked the biotinylated signaling probe to render the biosensor electrochemical response. The genosensor showed to reach a sensitivity of 174.4 μA fM and a limit of detection of 807 fM when using streptavidin poly-HRP20 enzymatic complex, detected SARS-CoV-2 specifically and discriminated it against homologous viruses in spiked samples and samples from SARS-CoV-2 cell cultures, a step forward to detect SARS-CoV-2 closer to the patient as a promising way for diagnosis and surveillance of COVID-19.
严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）感染可导致冠状病毒病（COVID-19）和当前的大流行。其死亡率增加，表明迫切需要急性和快速的诊断工具来替代当前的血清学检测和分子技术。电化学基因传感器设备的特点使其适合于更接近患者的快速和准确检测。本工作报道了一种用于 SARS-CoV-2 检测和区分同源呼吸道病毒的特定电化学基因传感器。电化学生物传感器是通过将巯基化捕获探针固定在马来酰亚胺涂覆的磁性颗粒上组装而成的，然后通过捕获和生物素化信号探针之间的夹心型特异性靶杂交来实现。这些探针经过严格的生物信息学设计和体外测试。基于链霉亲和素-辣根过氧化物酶的酶复合物将生物素化信号探针连接起来，赋予生物传感器电化学响应。当使用链霉亲和素多 HRP20 酶复合物时，该基因传感器的灵敏度达到 174.4 μA fM，检测限达到 807 fM，可特异性检测 SARS-CoV-2，并可区分在添加样本和 SARS-CoV-2 细胞培养物样本中的同源病毒，这是朝着更接近患者检测 SARS-CoV-2 的方向迈出的一步，是 COVID-19 诊断和监测的一种有前途的方法。
