Kinetic fingerprinting to identify and count single nucleic acids

Single unamplified miRNAs are detected with high specificity by kinetic fingerprinting. MicroRNAs (miRNAs) have emerged as promising diagnostic biomarkers. We introduce a kinetic fingerprinting approach called single-molecule recognition through equilibrium Poisson sampling (SiMREPS) for the amplifi...

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Bibliographic Details
Published in:Nature biotechnology 2015-07, Vol.33 (7), p.730-732
Main Authors: Johnson-Buck, Alexander, Su, Xin, Giraldez, Maria D, Zhao, Meiping, Tewari, Muneesh, Walter, Nils G
Format: Article
Language:English
Subjects:
631/61/32
639/638/11/874
639/638/11/876
Agriculture
Bioinformatics
Biological markers
Biomarkers
Biomedical Engineering/Biotechnology
Biomedicine
Biotechnology
brief-communication
Fluorescent Dyes - chemistry
Genetic research
Humans
Kinetics
Life Sciences
Limit of Detection
Methods
MicroRNA
MicroRNAs
MicroRNAs - analysis
MicroRNAs - chemistry
Nucleic acids
Polymorphism
Sequence Analysis, RNA - methods
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Summary:Single unamplified miRNAs are detected with high specificity by kinetic fingerprinting. MicroRNAs (miRNAs) have emerged as promising diagnostic biomarkers. We introduce a kinetic fingerprinting approach called single-molecule recognition through equilibrium Poisson sampling (SiMREPS) for the amplification-free counting of single unlabeled miRNA molecules, which circumvents thermodynamic limits of specificity and virtually eliminates false positives. We demonstrate high-confidence, single-molecule detection of synthetic and endogenous miRNAs in both buffer and minimally treated biofluids, as well as >500-fold discrimination between single nucleotide polymorphisms.
ISSN:1087-0156
1546-1696
DOI:10.1038/nbt.3246