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This work was financially supported by the Spanish Ministry of Economy and Competitiveness (PID2020-116728RB-I00, RED2022-134120-T, PID2023-150844OB-I00, PID2022-138908NB-C31, and PID2021-122299NB-I00), the Community of Madrid (REACT-UE NANOCOV-CM and Y2020/NMT6469), the Spanish Ministry of Science, Innovation and Universities (TED2021-129738B-I00, TED2021-130470B-I00, and TED2021-129999B-C32) and the "Maria de Maeztu" Programme for Units of Excellence in R&D, CEX2023-001316-M. Laura Gutierrez-Galvez gratefully acknowledges the financial support of a Formacion del Profesorado Universitario (FPU) grant from the Spanish Ministry of Universities (FPU19/06309). Estefania Enebral-Romero received financial support from the "Nanotecnologia para deteccion del SARS-CoV-2 y sus variantes, NANOCOV" project and a contract as a pre-doctoral researcher funded by a grant CEX2020-001039-S, supported by MCIN/AEI/10.13039/501100011033. We also acknowledge Maria U. Gonzalez and Raquel Alvaro for their help with fluorescence microscopy and EDAX mapping, respectively, and the services of the MiNa Laboratory at IMN and funding from CM (project S2018/NMT-4291 TEC2SPACE), MINECO (project CSIC13-4E-1794) and the EU (FEDER, FSE). We also acknowledge the "Servicio de Microbiologia, Hospital Universitario Ramon y Cajal and Instituto Ramon y Cajal de Investigacion Sanitaria (IRYCIS)" for providing the clinical samples. We acknowledge the support from the "(MAD2D-CM)-UAM" project funded by Comunidad de Madrid by the Recovery, Transformation and Resilience Plan and by NextGenerationEU from the European Union.

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Advancing diagnostics with BODIPY-bismuthene DNA biosensors

Publicated to:Nanoscale. 17 (13): 8126-8140 - 2025-03-06 17(13), DOI: 10.1039/d4nr05258g

Authors: Gutierrez-Galvez, Laura; Enebral-Romero, Estefania; Amores, Miguel Angel Valle; Coronado, Clara Pina; Torres, Inigo; Lopez-Diego, David; Luna, Monica; Fraile, Alberto; Zamora, Felix; Aleman, Jose; Alvarez, Jesus; Capitan, Maria Jose; Lorenzo, Encarnacion; Garcia-Mendiola, Tania

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Abstract

In this work, an electrochemical biosensor is prepared based on few-layer bismuthene hexagons (FLBHs) and a water-soluble BODIPY (BDP) derivative (BDP-NaSO3) for early infection diagnosis. In particular, the detection in advance of a virus sequence in nasopharyngeal swab samples was developed. The combination of the FLBHs and BDP-NaSO3 facilitates the direct, sensitive, and specific detection of gene viruses without the need for any prior amplification step. This work demonstrates that the FLBHs provide an improved electrochemical platform for immobilizing thiolated DNA capture probes that increase the sensitivity of the biosensor, while BDP-NaSO3 serves as a newly powerful electrochemical indicator of the hybridization event. As a proof of concept, SARS-CoV-2 was selected as the model virus. The developed biosensor demonstrated selective, rapid, and straightforward detection of the specific sequence RNA-dependent RNA-polymerase (RdRp) of SARS-CoV-2 with a detection limit of 4.97 fM and a linear range from 16.6 fM to 100 fM. Furthermore, this platform successfully detects the virus directly in nasopharyngeal swab samples with a viral load of at least 19 Cts without being subjected to any prior amplification stage. Finally, the high stability of the biosensor response, which has been working under ambient conditions for over one month, the selectivity and rapidity for specific virus detection, and the requirement of low-volume samples for the determination are remarkable characteristics that make it ideal for its potential application in clinical diagnosis in point-of-care settings.

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