Free PCR virus detection via few-layer bismuthene and tetrahedral DNA nanostructured assemblies

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 an...

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Detalhes bibliográficos
Autores: Gutiérrez-Gálvez, Laura, García-Fernández, Daniel, Barrio Redondo, Melisa del, Luna, Mónica, Torres, Íñigo, Zamora, Félix, Navío, Cristina, Milán-Rois, Paula, Castellanos, Milagros, Abreu, Melanie, Cantón, Rafael, Galán, Juan Carlos, Somoza, Álvaro, Miranda, Rodolfo, García-Mendiola, Tania, Lorenzo, Encarnación
Formato: artículo
Fecha de publicación:2024
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/362179
Acesso em linha:http://hdl.handle.net/10261/362179
https://api.elsevier.com/content/abstract/scopus_id/85177220280
Access Level:acceso abierto
Palavra-chave:Biocatalysis
DNA biosensor
Electrochemiluminescence
Few-layer bismuthene
SARS-CoV-2
Tetrahedral DNA nanostructures
Descrição
Resumo: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.