A Truncated Multi-Thiol Aptamer-Based SARS-CoV-2 Electrochemical Biosensor: Towards Variant-Specific Point-of-Care Detection with Optimized Fabrication
With the goal of fast and accurate diagnosis of infectious diseases, this study presents a novel electrochemical biosensor that employs a refined aptamer (C9t) for the detection of spike (S) protein SARS-CoV-2 variants in a flexible multielectrode aptasensor array with PoC capabilities. Two aptamer...
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MDPI AG
2025-01-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/15/1/24 |
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| author | Sergio Roberto Molina Ramirez Nafiseh Samiseresht Mateo Alejandro Martínez-Roque Ferdinando Catania Kevin Graef Martin Rabe Andreas Offenhäusser Dirk Mayer Gabriela Figueroa-Miranda |
| author_facet | Sergio Roberto Molina Ramirez Nafiseh Samiseresht Mateo Alejandro Martínez-Roque Ferdinando Catania Kevin Graef Martin Rabe Andreas Offenhäusser Dirk Mayer Gabriela Figueroa-Miranda |
| author_sort | Sergio Roberto Molina Ramirez |
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| description | With the goal of fast and accurate diagnosis of infectious diseases, this study presents a novel electrochemical biosensor that employs a refined aptamer (C9t) for the detection of spike (S) protein SARS-CoV-2 variants in a flexible multielectrode aptasensor array with PoC capabilities. Two aptamer modifications were employed: removing the primer binding sites and including two dithiol phosphoramidite anchor molecules. Thus, reducing fabrication time from 24 to 3 h and increasing the stability and sparseness for multi-thiol aptasensors compared to a standard aptasensor using single thiols, without a reduction in aptamer density. The biosensor fabrication, optimization, and detection were verified in detail by electrochemistry, QCM-D, SPR, and XPS. The analyte–receptor binding was further confirmed spectroscopically at the level of individual molecules by AFM-IR. The aptasensor possesses a low limit of detection (8.0 fg/mL), the highest sensitivity reported for S protein (209.5 signal per concentration decade), and a wide dynamic detection range (8.0 fg/mL–38 ng/mL) in nasopharyngeal samples, covering the clinically relevant range. Furthermore, the C9t aptasensor showed high selectivity for SARS-CoV-2 S proteins over biomarkers for MERS-CoV, RSV, and Influenza. Even more, it showed a three times higher sensitivity for the Omicron in comparison to the Wuhan strain (wild type), alpha, and beta variants of the SARS-CoV-2 virus. Those results demonstrate the creation of an affordable and variant-selective refined C9t aptasensor that outperformed current rapid diagnosis tests. |
| format | Article |
| id | doaj-art-fde362bc49b34de09a12fdf6f0843126 |
| institution | Kabale University |
| issn | 2079-6374 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj-art-fde362bc49b34de09a12fdf6f08431262025-01-24T13:25:27ZengMDPI AGBiosensors2079-63742025-01-011512410.3390/bios15010024A Truncated Multi-Thiol Aptamer-Based SARS-CoV-2 Electrochemical Biosensor: Towards Variant-Specific Point-of-Care Detection with Optimized FabricationSergio Roberto Molina Ramirez0Nafiseh Samiseresht1Mateo Alejandro Martínez-Roque2Ferdinando Catania3Kevin Graef4Martin Rabe5Andreas Offenhäusser6Dirk Mayer7Gabriela Figueroa-Miranda8Institute of Biological Information Processing, Bioelectronics (IBI-3), Forschungszentrum Jülich GmbH, 52428 Jülich, GermanyDepartment of Interface Chemistry and Surface Engineering, Max Planck Institute for Sustainable Materials GmbH, 40237 Düsseldorf, GermanyInstitute of Biological Information Processing, Bioelectronics (IBI-3), Forschungszentrum Jülich GmbH, 52428 Jülich, GermanyInstitute of Biological Information Processing, Bioelectronics (IBI-3), Forschungszentrum Jülich GmbH, 52428 Jülich, GermanyInstitute of Biological Information Processing, Bioelectronics (IBI-3), Forschungszentrum Jülich GmbH, 52428 Jülich, GermanyDepartment of Interface Chemistry and Surface Engineering, Max Planck Institute for Sustainable Materials GmbH, 40237 Düsseldorf, GermanyInstitute of Biological Information Processing, Bioelectronics (IBI-3), Forschungszentrum Jülich GmbH, 52428 Jülich, GermanyInstitute of Biological Information Processing, Bioelectronics (IBI-3), Forschungszentrum Jülich GmbH, 52428 Jülich, GermanyInstitute of Biological Information Processing, Bioelectronics (IBI-3), Forschungszentrum Jülich GmbH, 52428 Jülich, GermanyWith the goal of fast and accurate diagnosis of infectious diseases, this study presents a novel electrochemical biosensor that employs a refined aptamer (C9t) for the detection of spike (S) protein SARS-CoV-2 variants in a flexible multielectrode aptasensor array with PoC capabilities. Two aptamer modifications were employed: removing the primer binding sites and including two dithiol phosphoramidite anchor molecules. Thus, reducing fabrication time from 24 to 3 h and increasing the stability and sparseness for multi-thiol aptasensors compared to a standard aptasensor using single thiols, without a reduction in aptamer density. The biosensor fabrication, optimization, and detection were verified in detail by electrochemistry, QCM-D, SPR, and XPS. The analyte–receptor binding was further confirmed spectroscopically at the level of individual molecules by AFM-IR. The aptasensor possesses a low limit of detection (8.0 fg/mL), the highest sensitivity reported for S protein (209.5 signal per concentration decade), and a wide dynamic detection range (8.0 fg/mL–38 ng/mL) in nasopharyngeal samples, covering the clinically relevant range. Furthermore, the C9t aptasensor showed high selectivity for SARS-CoV-2 S proteins over biomarkers for MERS-CoV, RSV, and Influenza. Even more, it showed a three times higher sensitivity for the Omicron in comparison to the Wuhan strain (wild type), alpha, and beta variants of the SARS-CoV-2 virus. Those results demonstrate the creation of an affordable and variant-selective refined C9t aptasensor that outperformed current rapid diagnosis tests.https://www.mdpi.com/2079-6374/15/1/24electrochemical aptasensorSARS-CoV-2aptamer truncationmulti-thiolS protein |
| spellingShingle | Sergio Roberto Molina Ramirez Nafiseh Samiseresht Mateo Alejandro Martínez-Roque Ferdinando Catania Kevin Graef Martin Rabe Andreas Offenhäusser Dirk Mayer Gabriela Figueroa-Miranda A Truncated Multi-Thiol Aptamer-Based SARS-CoV-2 Electrochemical Biosensor: Towards Variant-Specific Point-of-Care Detection with Optimized Fabrication Biosensors electrochemical aptasensor SARS-CoV-2 aptamer truncation multi-thiol S protein |
| title | A Truncated Multi-Thiol Aptamer-Based SARS-CoV-2 Electrochemical Biosensor: Towards Variant-Specific Point-of-Care Detection with Optimized Fabrication |
| title_full | A Truncated Multi-Thiol Aptamer-Based SARS-CoV-2 Electrochemical Biosensor: Towards Variant-Specific Point-of-Care Detection with Optimized Fabrication |
| title_fullStr | A Truncated Multi-Thiol Aptamer-Based SARS-CoV-2 Electrochemical Biosensor: Towards Variant-Specific Point-of-Care Detection with Optimized Fabrication |
| title_full_unstemmed | A Truncated Multi-Thiol Aptamer-Based SARS-CoV-2 Electrochemical Biosensor: Towards Variant-Specific Point-of-Care Detection with Optimized Fabrication |
| title_short | A Truncated Multi-Thiol Aptamer-Based SARS-CoV-2 Electrochemical Biosensor: Towards Variant-Specific Point-of-Care Detection with Optimized Fabrication |
| title_sort | truncated multi thiol aptamer based sars cov 2 electrochemical biosensor towards variant specific point of care detection with optimized fabrication |
| topic | electrochemical aptasensor SARS-CoV-2 aptamer truncation multi-thiol S protein |
| url | https://www.mdpi.com/2079-6374/15/1/24 |
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