ArticleJournal of the American Chemical Society2018
Ultraspecific and Amplification-Free Quantification of Mutant DNA by Single-Molecule Kinetic Fingerprinting.
Article in Journal of the American Chemical Society, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed.
- FINDER converts zero-background kinetic fingerprinting into area-scalable attomolar biomarker detection.Research square · 2026Article
- FINDER converts zero-background kinetic fingerprinting into area-scalable attomolar biomarker detection.bioRxiv : the preprint server for biology · 2026Article
- Ultrafast and specific miRNA quantification via single-molecule fluorescence quenching kinetics.Communications biology · 2026Article
- Foundation model for efficient biological discovery in single-molecule time traces.Nature methods · 2025Article
- Chromato-Kinetic Fingerprinting Enables Multiomic Digital Counting of Single Disease Biomarker Molecules.ACS nano · 2025Article
- Analytical techniques for nucleic acid and protein detection with single-molecule sensitivity.Experimental & molecular medicine · 2025Review
- Chromato-kinetic fingerprinting enables multiomic digital counting of single disease biomarker molecules.bioRxiv : the preprint server for biology · 2025Article
- Ultrasensitive Amplification-Free Quantification of a Methyl CpG-Rich Cancer Biomarker by Single-Molecule Kinetic Fingerprinting.Analytical chemistry · 2024Article
- Deep learning based local feature classification to automatically identify single molecule fluorescence events.Communications biology · 2024Article
- Continuous Monitoring Biosensing Mediated by Single-Molecule Plasmon-Enhanced Fluorescence in Complex Matrices.ACS nano · 2024Article
- A mutation-sensitive, multiplexed and amplification-free detection of nucleic acids by stretching single-molecule tandem hairpin probes.Nucleic acids research · 2023Article
- Digital and Absolute Assays for Low Abundance Molecular Biomarkers.Accounts of chemical research · 2023Article
- Single-Molecule Optical Biosensing: Recent Advances and Future Challenges.ACS physical chemistry Au · 2023Review
- Massively Parallel Selection of NanoCluster Beacons.Advanced materials (Deerfield Beach, Fla.) · 2022Article
- Attomolar Sensitivity in Single Biomarker Counting upon Aqueous Two-Phase Surface Enrichment.ACS sensors · 2022Article
- Dynamic single-molecule sensing by actively tuning binding kinetics for ultrasensitive biomarker detection.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- A guide to accelerated direct digital counting of single nucleic acid molecules by FRET-based intramolecular kinetic fingerprinting.Methods (San Diego, Calif.) · 2022Review
- Rapid kinetic fingerprinting of single nucleic acid molecules by a FRET-based dynamic nanosensor.Biosensors & bioelectronics · 2021Article
- Recent developments in the characterization of nucleic acid hybridization kinetics.Current opinion in biomedical engineering · 2021Article
- DNA hybridisation kinetics using single-molecule fluorescence imaging.Essays in biochemistry · 2021Review
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
Conventional techniques for detecting rare DNA sequences require many cycles of PCR amplification for high sensitivity and specificity, potentially introducing significant biases and errors. While amplification-free methods exist, they rarely achieve the ability to detect single molecules, and their ability to discriminate between single-nucleotide variants is often dictated by the specificity limits of hybridization thermodynamics. Here we show that a direct detection approach using single-molecule kinetic fingerprinting can surpass the thermodynamic discrimination limit by 3 orders of magnitude, with a dynamic range of up to 5 orders of magnitude with optional super-resolution analysis. This approach detects mutations as subtle as the drug-resistance-conferring cancer mutation EGFR T790M (a single C → T substitution) with an estimated specificity of 99.99999%, surpassing even the leading PCR-based methods and enabling detection of 1 mutant molecule in a background of at least 1 million wild-type molecules. This level of specificity revealed rare, heat-induced cytosine deamination events that introduce false positives in PCR-based detection, but which can be overcome in our approach through milder thermal denaturation and enzymatic removal of damaged nucleobases.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.