title: Sensitive and multiplexed on-chip microRNA profiling in oil-isolated hydrogel chambers.
abstract: Although microRNAs (miRNAs) have been shown to be excellent indicators of disease state, current profiling platforms are insufficient for clinical translation. Here, we demonstrate a versatile hydrogel-based microfluidic approach and novel amplification scheme for entirely on-chip, sensitive, and highly specific miRNA detection without the risk of sequence bias. A simulation-driven approach is used to engineer the hydrogel geometry and the gel-reaction environment is chemically optimized for robust detection performance. The assay provides 22.6 fM sensitivity over a three log range, demonstrates multiplexing across at least four targets, and requires just 10.3 ng of total RNA input in a 2 hour and 15 minutes assay.
authors.affiliation: Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 (USA).
authors.forename: Hyewon
authors.initials: H
authors.lastname: Lee
authors.forename: Rathi L
authors.initials: RL
authors.lastname: Srinivas
authors.forename: Ankur
authors.initials: A
authors.lastname: Gupta
authors.forename: Patrick S
authors.initials: PS
authors.lastname: Doyle
mesh_terms: D020869:Gene Expression Profiling / Q000295:instrumentation*; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D035683:MicroRNAs / Q000032:analysis* / Q000378:metabolism; D046210:Microfluidic Analytical Techniques; D021141:Nucleic Acid Amplification Techniques; D009821:Oils / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis
chemical_list: D035683:MicroRNAs; D009821:Oils; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate
keywords: amplification; diagnostics; hydrogels; microfluidics