电化学发光
化学
检出限
线性范围
DNA
葡萄糖氧化酶
氧化酶试验
生物结合
组合化学
纳米技术
色谱法
生物传感器
生物化学
材料科学
酶
作者
Laura Gutiérrez-Gálvez,Daniel García-Fernández,Melisa del Barrio,María Virginia Luna,Íñigo Torres,Félix Zamora,Cristina Navío,Paula Milán-Rois,Milagros Castellanos,Melanie Abreu,Rafael Cantón,Juan Carlos Galán,Álvaro Somoza,Rodolfo Miranda,Tania García‐Mendiola,Encarnación Lorenzo
出处
期刊:Talanta
[Elsevier]
日期:2024-03-01
卷期号:269: 125405-125405
标识
DOI:10.1016/j.talanta.2023.125405
摘要
In this work we describe a highly sensitive method based on a biocatalyzed electrochemiluminescence approach. The system combines, for the first time, the use of few-layer bismuthene (FLB) as a platform for the oriented immobilization of tetrahedral DNA nanostructures (TDNs) specifically designed and synthetized to detect a specific SARS-CoV-2 gene sequence. In one of its vertices, these TDNs contain a DNA capture probe of the open reading frame 1 ab (ORF1ab) of the virus, available for the biorecognition of the target DNA/RNA. At the other three vertices, there are thiol groups that enable the stable anchoring/binding to the FLB surface. This novel geometry/approach enables not only the binding of the TDNs to surfaces, but also the orientation of the capture probe in a direction normal to the bismuthine surface so that it is readily accessible for binding/recognition of the specific SARS-CoV-2 sequence. The analytical signal is based on the anodic electrochemiluminescence (ECL) intensity of luminol which, in turn, arises as a result of the reaction with H2O2, generated by the enzymatic reaction of glucose oxidation, catalyzed by the biocatalytic label avidin-glucose oxidase conjugate (Av-GOx), which acts as co-reactant in the electrochemiluminescent reaction. The method exhibits a limit of detection (LOD) of 4.31 aM and a wide linear range from 14.4 aM to 1.00 μM, and its applicability was confirmed by detecting SARS-CoV-2 in nasopharyngeal samples from COVID-19 patients without the need of any amplification process.
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