Universal amplification-free molecular diagnostics by billion-fold hierarchical nanofluidic concentration
Rapid and reliable detection of ultralow-abundance nucleic acids and proteins in complex biological media may greatly advance clinical diagnostics and biotechnology development. Currently, nucleic acid tests rely on enzymatic processes for target amplification (e.g., PCR), which have many inherent i...
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| Format: | Article |
| Language: | English |
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Proceedings of the National Academy of Sciences
2020
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| Online Access: | https://hdl.handle.net/1721.1/124450 |
| _version_ | 1826199464995454976 |
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| author | Ouyang, Wei Han, Jongyoon |
| author2 | Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science |
| author_facet | Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Ouyang, Wei Han, Jongyoon |
| author_sort | Ouyang, Wei |
| collection | MIT |
| description | Rapid and reliable detection of ultralow-abundance nucleic acids and proteins in complex biological media may greatly advance clinical diagnostics and biotechnology development. Currently, nucleic acid tests rely on enzymatic processes for target amplification (e.g., PCR), which have many inherent issues restricting their implementation in diagnostics. On the other hand, there exist no protein amplification techniques, greatly limiting the development of protein-based diagnosis. We report a universal biomolecule enrichment technique termed hierarchical nanofluidic molecular enrichment system (HOLMES) for amplification-free molecular diagnostics using massively paralleled and hierarchically cascaded nanofluidic concentrators. HOLMES achieves billion-fold enrichment of both nucleic acids and proteins within 30 min, which not only overcomes many inherent issues of nucleic acid amplification but also provides unprecedented enrichment performance for protein analysis. HOLMES features the ability to selectively enrich target biomolecules and simultaneously deplete nontargets directly in complex crude samples, thereby enormously enhancing the signal-to-noise ratio of detection. We demonstrate the direct detection of attomolar nucleic acids in urine and serum within 35 min and HIV p24 protein in serum within 60 min. The performance of HOLMES is comparable to that of nucleic acid amplification tests and near million-fold improvement over standard enzyme-linked immunosorbent assay (ELISA) for protein detection, being much simpler and faster in both applications. We additionally measured human cardiac troponin I protein in 9 human plasma samples, and showed excellent agreement with ELISA and detection below the limit of ELISA. HOLMES is in an unparalleled position to unleash the potential of protein-based diagnosis. |
| first_indexed | 2024-09-23T11:20:39Z |
| format | Article |
| id | mit-1721.1/124450 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T11:20:39Z |
| publishDate | 2020 |
| publisher | Proceedings of the National Academy of Sciences |
| record_format | dspace |
| spelling | mit-1721.1/1244502022-09-27T18:52:57Z Universal amplification-free molecular diagnostics by billion-fold hierarchical nanofluidic concentration Ouyang, Wei Han, Jongyoon Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Massachusetts Institute of Technology. Research Laboratory of Electronics Massachusetts Institute of Technology. Department of Biological Engineering Multidisciplinary Rapid and reliable detection of ultralow-abundance nucleic acids and proteins in complex biological media may greatly advance clinical diagnostics and biotechnology development. Currently, nucleic acid tests rely on enzymatic processes for target amplification (e.g., PCR), which have many inherent issues restricting their implementation in diagnostics. On the other hand, there exist no protein amplification techniques, greatly limiting the development of protein-based diagnosis. We report a universal biomolecule enrichment technique termed hierarchical nanofluidic molecular enrichment system (HOLMES) for amplification-free molecular diagnostics using massively paralleled and hierarchically cascaded nanofluidic concentrators. HOLMES achieves billion-fold enrichment of both nucleic acids and proteins within 30 min, which not only overcomes many inherent issues of nucleic acid amplification but also provides unprecedented enrichment performance for protein analysis. HOLMES features the ability to selectively enrich target biomolecules and simultaneously deplete nontargets directly in complex crude samples, thereby enormously enhancing the signal-to-noise ratio of detection. We demonstrate the direct detection of attomolar nucleic acids in urine and serum within 35 min and HIV p24 protein in serum within 60 min. The performance of HOLMES is comparable to that of nucleic acid amplification tests and near million-fold improvement over standard enzyme-linked immunosorbent assay (ELISA) for protein detection, being much simpler and faster in both applications. We additionally measured human cardiac troponin I protein in 9 human plasma samples, and showed excellent agreement with ELISA and detection below the limit of ELISA. HOLMES is in an unparalleled position to unleash the potential of protein-based diagnosis. National Institutes of Health (U.S.) (Grant U19AI10975) National Institutes of Health (U.S.) (Grant R01AI117043) 2020-03-31T14:53:01Z 2020-03-31T14:53:01Z 2019-07-29 2020-02-13T13:30:19Z Article http://purl.org/eprint/type/JournalArticle 0027-8424 1091-6490 https://hdl.handle.net/1721.1/124450 Ouyang, Wei and Jongyoon Han. "Universal amplification-free molecular diagnostics by billion-fold hierarchical nanofluidic concentration." Proceedings of the National Academy of Sciences of the United States of America 116 (2019): 16240-16249 © 2019 The Author(s) en 10.1073/pnas.1904513116 Proceedings of the National Academy of Sciences of the United States of America Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Proceedings of the National Academy of Sciences PNAS |
| spellingShingle | Multidisciplinary Ouyang, Wei Han, Jongyoon Universal amplification-free molecular diagnostics by billion-fold hierarchical nanofluidic concentration |
| title | Universal amplification-free molecular diagnostics by billion-fold hierarchical nanofluidic concentration |
| title_full | Universal amplification-free molecular diagnostics by billion-fold hierarchical nanofluidic concentration |
| title_fullStr | Universal amplification-free molecular diagnostics by billion-fold hierarchical nanofluidic concentration |
| title_full_unstemmed | Universal amplification-free molecular diagnostics by billion-fold hierarchical nanofluidic concentration |
| title_short | Universal amplification-free molecular diagnostics by billion-fold hierarchical nanofluidic concentration |
| title_sort | universal amplification free molecular diagnostics by billion fold hierarchical nanofluidic concentration |
| topic | Multidisciplinary |
| url | https://hdl.handle.net/1721.1/124450 |
| work_keys_str_mv | AT ouyangwei universalamplificationfreemoleculardiagnosticsbybillionfoldhierarchicalnanofluidicconcentration AT hanjongyoon universalamplificationfreemoleculardiagnosticsbybillionfoldhierarchicalnanofluidicconcentration |