ArticleNucleic acids research2017
Sensitive and inexpensive digital DNA analysis by microfluidic enrichment of rolling circle amplified single-molecules.
Article in Nucleic acids research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Rolling Circle Amplification as a Molecular Tool for Spatially Resolved Signal Amplification in Single Molecule Counting Assay.Biosensors · 2025Review
- Digital and Absolute Assays for Low Abundance Molecular Biomarkers.Accounts of chemical research · 2023Article
- Cell-Free Production Systems in Droplet Microfluidics.Advances in biochemical engineering/biotechnology · 2023Review
- Rolling Circle Amplification Tailored for Plasmonic Biosensors: From Ensemble to Single-Molecule Detection.ACS applied materials & interfaces · 2022Article
- Rolling Circle Amplification in Integrated Microsystems: An Uncut Gem toward Massively Multiplexed Pathogen Diagnostics and Genotyping.Accounts of chemical research · 2021Review
- A 3D-Printed Microfluidic Device for qPCR Detection of Macrolide-Resistant Mutations ofBiosensors · 2021Article
- Bead-based Padlock Rolling Circle Amplification under Molecular Crowding Conditions: The Effects of Crowder Charge and Size.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2021Article
- Recent development of antibiotic detection in food and environment: the combination of sensors and nanomaterials.Mikrochimica acta · 2021Review
- Directed co-evolution of interacting protein-peptide pairs by compartmentalized two-hybrid replication (C2HR).Nucleic acids research · 2020Article
- A multiplex platform for digital measurement of circular DNA reaction products.Nucleic acids research · 2020Article
- Ultrasensitive Detection of Attomolar Protein Concentrations by Dropcast Single Molecule Assays.Journal of the American Chemical Society · 2020Article
- Characterization of Binding of Magnetic Nanoparticles to Rolling Circle Amplification Products by Turn-On Magnetic Assay.Biosensors · 2019Article
- A novel mutation tolerant padlock probe design for multiplexed detection of hypervariable RNA viruses.Scientific reports · 2019Article
- Development of a Sensitive Induction-Based Magnetic Nanoparticle Biodetection Method.Nanomaterials (Basel, Switzerland) · 2018Article
- Interlinked DNA nano-circles for measuring topoisomerase II activity at the level of single decatenation events.Nucleic acids research · 2017Article
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
Single molecule quantification assays provide the ultimate sensitivity and precision for molecular analysis. However, most digital analysis techniques, i.e. droplet PCR, require sophisticated and expensive instrumentation for molecule compartmentalization, amplification and analysis. Rolling circle amplification (RCA) provides a simpler means for digital analysis. Nevertheless, the sensitivity of RCA assays has until now been limited by inefficient detection methods. We have developed a simple microfluidic strategy for enrichment of RCA products into a single field of view of a low magnification fluorescent sensor, enabling ultra-sensitive digital quantification of nucleic acids over a dynamic range from 1.2 aM to 190 fM. We prove the broad applicability of our analysis platform by demonstrating 5-plex detection of as little as ∼1 pg (∼300 genome copies) of pathogenic DNA with simultaneous antibiotic resistance marker detection, and the analysis of rare oncogene mutations. Our method is simpler, more cost-effective and faster than other digital analysis techniques and provides the means to implement digital analysis in any laboratory equipped with a standard fluorescent microscope.
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