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Updated: Jul 12, 2026
High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
Published on: August 3, 2011
Compartmentalization-inspired dual-chamber CRISPR sensing coupled with single-atom electrocatalysis for
Jiangbo Dong1, Xinyao Li2, Tao Gu2
1Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, China; Department of Neurosurgery, Laboratory of Neurological Diseases and Interdisciplinary Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
This study presents a bioinspired dual-chamber sensing platform for multiplex microRNA (miRNA) analysis. The innovative design minimizes crosstalk, enabling sensitive and accurate detection of miRNAs for diagnostic applications.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Biosensing
Background:
- Cellular compartmentalization enhances biochemical process fidelity.
- Multiplex microRNA (miRNA) analysis is crucial for diagnostics but often suffers from crosstalk.
- Existing methods require complex procedures and lack sensitivity.
Purpose of the Study:
- To develop a compartmentalization-inspired platform for multiplex miRNA analysis with minimized crosstalk.
- To achieve high sensitivity and specificity in simultaneous miRNA detection.
- To validate the platform's applicability in biological samples.
Main Methods:
- A dual-chamber sensing platform utilizing padlock-probe ligation and rolling-circle-extension-driven loop-mediated isothermal amplification (R-LAMP).
- CRISPR/Cas12a system for specific amplicon activation and DNA biogate cleavage.
- Release of electroactive reporters (TMB, MB) from Fe-MOF nanocontainers.
- Electrochemical readout using a screen-printed electrode with a Co-N-C single-atom catalyst.
Main Results:
- Femtomolar detection limits (0.87 fM for miRNA-21, 0.72 fM for miRNA-155) in simultaneous assays.
- Broad linear range (1 fM-100 pM) and high discrimination against mismatched sequences.
- Accurate miRNA quantification in diluted human serum and cell lysates (validated by RT-qPCR).
Conclusions:
- The developed platform effectively minimizes crosstalk for multiplex miRNA analysis.
- Integration of bioinspired compartmentalization, CRISPR, and single-atom electrocatalysis offers a generalizable diagnostic approach.
- The platform demonstrates high sensitivity, specificity, and practical applicability for nucleic acid diagnostics.
