High-sensitivity, protein-independent detection of dsDNA sequences

Current methodologies for detecting the sequence of double-stranded DNA (dsDNA) require amplifying and denaturing the target into single-stranded DNA (ssDNA) to enable sequence detection through Watson-Crick base pairing. However, these approaches are limited by the risks of nonspecific amplification, reliance on complex, temperature-sensitive protein enzymes, and harsh reaction conditions, such as in strong base or acidic environments. Here, we introduce a dsDNA detection platform that integrates a peptide nucleic acid (PNA) as the dsDNA denaturation agent, with multicomponent deoxyribozyme as the ssDNA detection tool, in a droplet-based system. This protein- and amplification-free method offers single-nucleotide resolution, detects down to a single dsDNA molecule, and delivers results within 1 h at room temperature. This work introduces a conceptually unique approach, that may be useful for both diagnostics and therapeutics.

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Publication Details

Journal
Proceedings of the National Academy of Sciences
Published
2026-02-04
DOI
https://doi.org/10.1073/pnas.2515765123
Citations
1
Primary Topic
Innovative Microfluidic and Catalytic Techniques Innovation
Type
article
Field-Weighted Citation Impact
21.28

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High-sensitivity, protein-independent detection of dsDNA sequences

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High-sensitivity, protein-independent detection of dsDNA sequences

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article en
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Abstract

Current methodologies for detecting the sequence of double-stranded DNA (dsDNA) require amplifying and denaturing the target into single-stranded DNA (ssDNA) to enable sequence detection through Watson-Crick base pairing. However, these approaches are limited by the risks of nonspecific amplification, reliance on complex, temperature-sensitive protein enzymes, and harsh reaction conditions, such as in strong base or acidic environments. Here, we introduce a dsDNA detection platform that integrates a peptide nucleic acid (PNA) as the dsDNA denaturation agent, with multicomponent deoxyribozyme as the ssDNA detection tool, in a droplet-based system. This protein- and amplification-free method offers single-nucleotide resolution, detects down to a single dsDNA molecule, and delivers results within 1 h at room temperature. This work introduces a conceptually unique approach, that may be useful for both diagnostics and therapeutics.

Proceedings of the National Academy of SciencesVol. 123(6)
Åbo Akademi University (FI), Harvard University (US), University of Turku (FI), Turku Centre for Computer Science (FI), Wenzhou Medical University (CN), First Affiliated Hospital of Wenzhou Medical University (CN), Heilbronn University (DE), Aalto University (FI)
Alexander von Humboldt-Stiftung, Harvard University, University of Pittsburgh, National Natural Science Foundation of China, National Institutes of Health, Natural Science Foundation of Zhejiang Province
Openalex Percentile: Top 1%
Innovative Microfluidic and Catalytic Techniques Innovation
21.28
DOIOpen Access PDFOpenAlex
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