Dendrimer-based DNA biosensor for HIV virus detection

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dc.contributor.affiliation000000041762408X
dc.contributor.affiliationDepartamento de Química Analítica y Análisis Instrumental, Universidad Autónoma de Madrid, Madrid, Spain; IMDEA Nanociencia, Madrid, Spain; CSIC - Instituto de Micro y Nanotecnología (IMN-CNM), Tres Cantos, Spain; Department of Molecular Biology, CSIC-UAM - Centro de Biología Molecular Severo Ochoa (CBM), Madrid, Spain; IFIMAC-Condensed Matter Physics Center, Madrid, Spain; Universidad Autónoma de Madrid, Madrid, Spain; Clínicas, Hospital Universitario La Paz, Madrid, Spain; Unidad de VIH, Hospital Universitario La Paz, Madrid, Spainen
dc.contributor.authorEnebral-Romero, E.
dc.contributor.authorToldos-Torres, M.
dc.contributor.authorLópez-Diego, D.
dc.contributor.authorLuna, M.
dc.contributor.authorFailde, M.
dc.contributor.authorGonzález-Tobío, B.
dc.contributor.authorZamora, F.
dc.contributor.authorFalces, I.
dc.contributor.authorMontes, M.L.
dc.contributor.authorGarcía-Mendiola, T.
dc.date.accessioned2025-10-30T16:14:32Z
dc.date.available2025-10-30T16:14:32Z
dc.date.issued2025
dc.format.mimetypeapplication/pdfen
dc.identifier.doi10.1016/j.bios.2025.118014en
dc.identifier.urihttps://hdl.handle.net/20.500.12614/4110
dc.issue.number118014en
dc.journal.titleBiosensors and Bioelectronicsen
dc.language.isoenen
dc.relation.projectIDTEMPFunding text 1: The synthesis of the HIV-based DNA dendrimer follows a divergent-growth strategy from Y-shaped DNA monomers (YA, YB, and YC) through the hybridization of the complementary nitrogenous bases. YA (red Y DNA monomer) and YB (blue Y DNA monomer) are composed of three 40-base DNA sequences each (TriX-1, TriX-2 and TriX-3), while YC includes two 61-base (TriC-1 and TriC-2) and one 40-base DNA sequence. Each monomer contains 12-base sticky-end fragments at the 5\u2032-end, enabling sequential hybridization and the stepwise formation of the dendrimer generations (G0 to G2). The TriC-1 and TriC-2 sequences contain two 5\u2032-thiolated 24-base sequences that correspond to the specific HIV probe sequence (yellow sequences), which remain free and available for the hybridization reaction with the target analyte. All sequences are listed and highlighted in different colours in Table S1 of SI and shown in Scheme 1B. Monomers were assembled by mixing equimolar amounts (1.00 \u03BCM) of each strand in TE buffer, and dendrimer formation was achieved by mixing YA, YB, and YC in a 1:3:6 ratio. Hybridizations were performed in a thermocycler using temperature-controlled steps, with melting temperatures (Tm) calculated to prevent non-specific binding. All details regarding the synthesis of each monomer and the complete structure of the HIV-based DNA dendrimer are provided in the Experimental and Results sections of the Supporting Information.Additional complementary characterization studies were also carried out using fluorescence microscopy, Raman spectroscopy, and EIS (Fig. S3, which are discussed in detail in Section 3.3 of the Supporting Information).To demonstrate the improvements in sensitivity and amplification capacity of the HIV-based DNA dendrimer, a calibration was performed using the CSPE/FLB/HIV-SH platform (see Scheme S1 in the Supporting Information), for concentrations of the HIVc sequence ranging from 500 fM to 100 pM. Fig. S5 in the SI presents the calibration plot obtained for the normalized current intensity registered (calculated as previously described) versus the logarithm of the HIVc concentration. The linear equation describes the upward linear correlation observed: |(i-iB)/iB| = 0.238\u00B7log[HIVc] + 0.218 (R = 0.9944) (n = 3). These results yield a sensitivity of 0.238 a.u. log (pM-1) and a CV of 2.27 %. The detection limit obtained was 0.235 pM (calculated using the equation y = XB + 3\u00B7SB, as explained previously). Comparing this with the LOD obtained for the HIV dendrimer-based DNA biosensor, it can be affirmed that the sensitivity of the device improves considerably, further demonstrating the benefits that the use of DNA dendrimers brings to biosensor design.This work was financially supported by the Spanish Ministry of Science, Innovation and Universities through the following projects: MCIU-25-RED2024-153970-T, PID2023-150844OB-I00, RED2022-134120-T, PID2022-138908NB-C31, PDC2021-120782-C21, and TED2021-129738B-I00. Estefan\u00EDa Enebral-Romero acknowledges funding from the project \u201CNanotecnolog\u00EDa para detecci\u00F3n del SARS-CoV-2 y sus variantes (NANOCOV)\u201D and a predoctoral research contract supported by grant CEX2020-001039-S, funded by MCIN/AEI/10.13039/501100011033. F.Z. acknowledges support from Spanish Ministry of Science, Innovation and Universities with the grant PID2022-138908NB-C31, from the Community of Madrid, the European Union's NextGenerationEU programme and the Recovery, Transformation and Resilience Plan, with the \u201CMAD2D-CM-UAM\u201D project, and from the Spanish Ministry of Science and Innovation through the \u201CMar\u00EDa de Maeztu\u201D Programme for Units of Excellence in R&D (CEX2023-001316-M). B. G. acknowledges the financial support from the \u201CAyuda para Contratos Predoctorales para la Formaci\u00F3n de Doctores\u201D program of MICINN (PRE2020-093162). Additional support was provided by the European Innovation Council under grant agreement 101047081 (EVA). We also acknowledge the MiNa Laboratory at IMN for technical support, and funding from the MCIN/AEI/10.13039/501100011033, EU (FEDER, FSE), and Next Generation EU/PRTR (EQC2021-006944-P). Special thanks are extended to Iker Falces and Mar\u00EDa Luisa Montes from Hospital Universitario La Paz for their collaboration in providing clinical plasma samples from HIV patients.; Funding text 2: F.Z. acknowledges support from Spanish Ministry of Science, Innovation and Universities with the grant PID2022-138908NB-C31, from the Community of Madrid , the European Union's NextGenerationEU programme and the Recovery, Transformation and Resilience Plan, with the \u201CMAD2D-CM-UAM\u201D project, and from the Spanish Ministry of Science and Innovation through the \u201CMar\u00EDa de Maeztu\u201D Programme for Units of Excellence in R&D (CEX2023-001316-M). B. G. acknowledges the financial support from the \u201CAyuda para Contratos Predoctorales para la Formaci\u00F3n de Doctores\u201D program of MICINN (PRE2020-093162). Additional support was provided by the European Innovation Council under grant agreement 101047081 (EVA).; Funding text 3: This work was financially supported by the Spanish Ministry of Science, Innovation and Universities through the following projects: MCIU-25-RED2024-153970-T, PID2023-150844OB-I00, RED2022-134120-T, PID2022-138908NB-C31, PDC2021-120782-C21, and TED2021-129738B-I00. Estefan\u00EDa Enebral-Romero acknowledges funding from the project \u201CNanotecnolog\u00EDa para detecci\u00F3n del SARS-CoV-2 y sus variantes (NANOCOV)\u201D and a predoctoral research contract supported by grant CEX2020-001039-S, funded by MCIN / AEI /10.13039/501100011033.; Funding text 4: We also acknowledge the MiNa Laboratory at IMN for technical support, and funding from the MCIN/AEI/10.13039/501100011033, EU (FEDER, FSE), and Next Generation EU/PRTR (EQC2021-006944-P). Special thanks are extended to Iker Falces and Mar\u00EDa Luisa Montes from Hospital Universitario La Paz for their collaboration in providing clinical plasma samples from HIV patients.
dc.rightsAtribución-NoComercial-SinDerivadas 3.0 España
dc.rights.accessRightsopen accessen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/es/
dc.titleDendrimer-based DNA biosensor for HIV virus detectionen
dc.typeresearch articleen
dc.type.hasVersionVoRen
dc.volume.number290en

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