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Updated: Oct 29, 2026
High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
Published on: August 3, 2011
Functional double-emulsion droplets enable multiplexed CRISPR quantification of cancer-related miRNAs
Yang Zhang1, Fei Deng2, Rui Sang2
1School of Mechanical and Manufacturing Engineering, Faculty of Engineering, University of New South Wales, Sydney, NSW, 2052, Australia.
Abstract:
Accurate, multiplexed microRNA detection is critical for early cancer diagnosis. We present a droplet-based digital CRISPR/Cas13a platform for simultaneous, amplification-free detection of multiple miRNAs. By integrating double emulsion (DE) microfluidics with programmable crRNAs, the system achieves orthogonal, interference-free readouts, enabling precise absolute quantification in complex biological samples. The assay demonstrates high sequence specificity, robust orthogonality, and consistent performance across target combinations and matrices. Fluorescence-positive droplet counts correlate linearly with input concentrations, allowing quantification of miRNA-21, miRNA-29a, and miRNA-92a with 10 copies/μL representing the lowest concentration experimentally evaluated in this study. The applicability of the platform is further evaluated in a small proof-of-concept cohort of plasma samples from healthy controls and colorectal cancer (CRC) patients. This platform provides an amplification-free approach for multiplexed miRNA quantification and supports further development for clinical biomarker analysis.
