Enhancing Colorimetric Detection of Nucleic Acids on Nitrocellulose Membranes: Cutting-Edge Applications in Diagnostics and Forensics
This study re-introduces a protein-free rapid test method for nucleic acids on paper based lateral flow assays utilizing special multichannel nitrocellulose membranes and DNA-Gold conjugates, achieving significantly enhanced sensitivity, easier protocols, reduced time of detection, reduced costs of...
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MDPI AG
2024-09-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/14/9/430 |
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| author | Nidhi Subhashini Yannick Kerler Marcus M. Menger Olga Böhm Judith Witte Christian Stadler Alexander Griberman |
| author_facet | Nidhi Subhashini Yannick Kerler Marcus M. Menger Olga Böhm Judith Witte Christian Stadler Alexander Griberman |
| author_sort | Nidhi Subhashini |
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| description | This study re-introduces a protein-free rapid test method for nucleic acids on paper based lateral flow assays utilizing special multichannel nitrocellulose membranes and DNA-Gold conjugates, achieving significantly enhanced sensitivity, easier protocols, reduced time of detection, reduced costs of production and advanced multiplexing possibilities. A protein-free nucleic acid-based lateral flow assay (NALFA) with a limit of detection of 1 pmol of DNA is shown for the first time. The total production duration of such an assay was successfully reduced from the currently known several days to just a few hours. The simplification and acceleration of the protocol make the method more accessible and practical for various applications. The developed method supports multiplexing, enabling the simultaneous detection of up to six DNA targets. This multiplexing capability is a significant improvement over traditional line tests and offers more comprehensive diagnostic potential in a single assay. The approach significantly reduces the run time compared to traditional line tests, which enhances the efficiency of diagnostic procedures. The protein-free aspect of this assay minimizes the prevalent complications of cross-reactivity in immunoassays especially in cases of multiplexing. It is also demonstrated that the NALFA developed in this study is amplification-free and hence does not rely on specialized technicians, nor does it involve labour-intensive steps like DNA extraction and PCR processes. Overall, this study presents a robust, efficient, and highly sensitive platform for DNA or RNA detection, addressing several limitations of current methods documented in the literature. The advancements in sensitivity, cost reduction, production time, and multiplexing capabilities mark a substantial improvement, holding great potential for various applications in diagnostics, forensics, and molecular biology. |
| format | Article |
| id | doaj-art-ffcbce1dd0654268bc8a4e505b6e8ddf |
| institution | OA Journals |
| issn | 2079-6374 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | MDPI AG |
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| series | Biosensors |
| spelling | doaj-art-ffcbce1dd0654268bc8a4e505b6e8ddf2025-08-20T01:56:11ZengMDPI AGBiosensors2079-63742024-09-0114943010.3390/bios14090430Enhancing Colorimetric Detection of Nucleic Acids on Nitrocellulose Membranes: Cutting-Edge Applications in Diagnostics and ForensicsNidhi Subhashini0Yannick Kerler1Marcus M. Menger2Olga Böhm3Judith Witte4Christian Stadler5Alexander Griberman6SERATEC Gesellschaft für Biotechnologie mbH, Ernst-Ruhstrat-Str. 5, 37079 Goettingen, GermanyFraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses (IZI-BB), Am Mühlenberg 13, 14476 Potsdam, GermanyFraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses (IZI-BB), Am Mühlenberg 13, 14476 Potsdam, GermanySartorius Stedim Biotech GmbH, August-Spindler-Str. 11, 37079 Goettingen, GermanySartorius Stedim Biotech GmbH, August-Spindler-Str. 11, 37079 Goettingen, GermanySERATEC Gesellschaft für Biotechnologie mbH, Ernst-Ruhstrat-Str. 5, 37079 Goettingen, GermanySERATEC Gesellschaft für Biotechnologie mbH, Ernst-Ruhstrat-Str. 5, 37079 Goettingen, GermanyThis study re-introduces a protein-free rapid test method for nucleic acids on paper based lateral flow assays utilizing special multichannel nitrocellulose membranes and DNA-Gold conjugates, achieving significantly enhanced sensitivity, easier protocols, reduced time of detection, reduced costs of production and advanced multiplexing possibilities. A protein-free nucleic acid-based lateral flow assay (NALFA) with a limit of detection of 1 pmol of DNA is shown for the first time. The total production duration of such an assay was successfully reduced from the currently known several days to just a few hours. The simplification and acceleration of the protocol make the method more accessible and practical for various applications. The developed method supports multiplexing, enabling the simultaneous detection of up to six DNA targets. This multiplexing capability is a significant improvement over traditional line tests and offers more comprehensive diagnostic potential in a single assay. The approach significantly reduces the run time compared to traditional line tests, which enhances the efficiency of diagnostic procedures. The protein-free aspect of this assay minimizes the prevalent complications of cross-reactivity in immunoassays especially in cases of multiplexing. It is also demonstrated that the NALFA developed in this study is amplification-free and hence does not rely on specialized technicians, nor does it involve labour-intensive steps like DNA extraction and PCR processes. Overall, this study presents a robust, efficient, and highly sensitive platform for DNA or RNA detection, addressing several limitations of current methods documented in the literature. The advancements in sensitivity, cost reduction, production time, and multiplexing capabilities mark a substantial improvement, holding great potential for various applications in diagnostics, forensics, and molecular biology.https://www.mdpi.com/2079-6374/14/9/430lateral flow assaymultiplexingnucleic acid lateral flow assay (NALFA)molecular beaconprotein-freeamplification-free |
| spellingShingle | Nidhi Subhashini Yannick Kerler Marcus M. Menger Olga Böhm Judith Witte Christian Stadler Alexander Griberman Enhancing Colorimetric Detection of Nucleic Acids on Nitrocellulose Membranes: Cutting-Edge Applications in Diagnostics and Forensics Biosensors lateral flow assay multiplexing nucleic acid lateral flow assay (NALFA) molecular beacon protein-free amplification-free |
| title | Enhancing Colorimetric Detection of Nucleic Acids on Nitrocellulose Membranes: Cutting-Edge Applications in Diagnostics and Forensics |
| title_full | Enhancing Colorimetric Detection of Nucleic Acids on Nitrocellulose Membranes: Cutting-Edge Applications in Diagnostics and Forensics |
| title_fullStr | Enhancing Colorimetric Detection of Nucleic Acids on Nitrocellulose Membranes: Cutting-Edge Applications in Diagnostics and Forensics |
| title_full_unstemmed | Enhancing Colorimetric Detection of Nucleic Acids on Nitrocellulose Membranes: Cutting-Edge Applications in Diagnostics and Forensics |
| title_short | Enhancing Colorimetric Detection of Nucleic Acids on Nitrocellulose Membranes: Cutting-Edge Applications in Diagnostics and Forensics |
| title_sort | enhancing colorimetric detection of nucleic acids on nitrocellulose membranes cutting edge applications in diagnostics and forensics |
| topic | lateral flow assay multiplexing nucleic acid lateral flow assay (NALFA) molecular beacon protein-free amplification-free |
| url | https://www.mdpi.com/2079-6374/14/9/430 |
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