title: Kinetic fingerprinting to identify and count single nucleic acids.
abstract: 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.
authors.affiliation: Single Molecule Analysis Group, Department of Chemistry, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Johnson-Buck
authors.affiliation: Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Su
authors.affiliation: Department of Internal Medicine, Division of Hematology/Oncology, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Maria D
authors.initials: MD
authors.lastname: Giraldez
authors.affiliation: Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing, China.
authors.forename: Meiping
authors.initials: M
authors.lastname: Zhao
authors.affiliation: 1] Department of Internal Medicine, Division of Hematology/Oncology, University of Michigan, Ann Arbor, Michigan, USA. [2] Department of Internal Medicine, Division of Molecular Medicine and Genetics, University of Michigan, Ann Arbor, Michigan, USA. [3] Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, USA. [4] Center for Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, Michigan, USA. [5] Biointerfaces Institute, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Muneesh
authors.initials: M
authors.lastname: Tewari
authors.affiliation: Single Molecule Analysis Group, Department of Chemistry, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Nils G
authors.initials: NG
authors.lastname: Walter
mesh_terms: D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D057230:Limit of Detection; D035683:MicroRNAs / Q000032:analysis* / Q000737:chemistry; D017423:Sequence Analysis, RNA / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D035683:MicroRNAs