Grupo de Sensores Químicos y Biosensores, Departamento de Química Analítica y Análisis Instrumental, Universidad Autónoma de Madrid, 28049, Madrid, Spain; Institute for Advanced Research in Chemical Sciences (IAdChem), Universidad Autónoma de Madrid, 28049, Madrid, Spain.
Grupo de Sensores Químicos y Biosensores, Departamento de Química Analítica y Análisis Instrumental, Universidad Autónoma de Madrid, 28049, Madrid, Spain.
Talanta. 2024 Dec 1;280:126708. doi: 10.1016/j.talanta.2024.126708. Epub 2024 Aug 13.
The development of rapid, accurate, sensitive, and low-cost diagnostic methods for COVID-19 detection in real-time is the unique way to control infection sources and monitor illness progression. In this work, we propose an electrochemical biosensor for the rapid and accuracy diagnosis of COVID-19, through the determination of ORF specific sequence. The biosensor is based on the immobilization of a thiolated sequence partially complementary (domain 1) to ORF on gold screen-printed electrodes and the use of bifunctional Au@Pt/Au core@shell nanoparticles modified with a second thiolated sequence partially complementary to ORF (domain 2) as electrochemical indicator of the hybridization of DNA sequences. The synthesized Au@Pt/Au nanoparticles consist of an Au core, a shell of Pt (Au@Pt NPs), that provides an excellent electrocatalytic activity toward the oxygen reduction reaction (ORR) even after formation of hybrid biomaterials by modification, through the Au protuberances growth on the NPs surface, with an oligonucleotide with recognition ability. The ORR electrochemical activity, enhanced by the label element (Au@Pt/Au NPs), has been employed, for the first time, as indicator of the hybridization event. Based on this strategy, target sequences of the SARS-CoV-2 virus have been detected with a detection limit of 32 pM. The selectivity of the biosensor was confirmed by analysing ORF sequence in the presence of DNA sequences from other viruses. The biosensor has been successfully applied to the direct detection of the virus in non-amplified samples of nasopharyngeal swabs from infected and non-infected patients. Results compare well with those obtained through RT-qPCR but our method is more rapid since does not need any amplification process.
开发快速、准确、灵敏且低成本的实时 COVID-19 诊断方法是控制传染源和监测疾病进展的独特方法。在这项工作中,我们提出了一种基于电化学的生物传感器,用于快速准确地诊断 COVID-19,通过确定 ORF 特定序列。该生物传感器基于将部分互补(域 1)到 ORF 的硫醇化序列固定在金丝网印刷电极上,以及使用带有第二个部分互补到 ORF(域 2)的硫醇化序列的双功能 Au@Pt/Au 核@壳纳米粒子作为 DNA 序列杂交的电化学指示剂。合成的 Au@Pt/Au 纳米粒子由 Au 核和 Pt 壳(Au@Pt NPs)组成,即使在通过 NPs 表面上的 Au 突起生长修饰形成杂交生物材料后,也对氧还原反应(ORR)具有极好的电催化活性,带有识别能力的寡核苷酸。通过标记元素(Au@Pt/Au NPs)增强的 ORR 电化学活性,首次被用作杂交事件的指示剂。基于这一策略,已经可以检测到 SARS-CoV-2 病毒的靶序列,检测限为 32 pM。通过分析来自其他病毒的 DNA 序列存在下的 ORF 序列,确认了生物传感器的选择性。该生物传感器已成功应用于直接检测来自感染和未感染患者的鼻咽拭子未经扩增的样本中的病毒。与通过 RT-qPCR 获得的结果相比,该方法的检测限为 32 pM。结果相当,但我们的方法更快,因为它不需要任何扩增过程。
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