Sensitive and Multiplexed On-chip microRNA Profiling in Oil-Isolated Hydrogel Chambers

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 22.6fM sensitivity over a three log range, demonstrates multiplexing across at least four targets, and requires just 10.3ng of total RNA input in a 2hour and 15minutes assay.