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    Area of Science:

    • Nanotechnology
    • Biosensing
    • Analytical Chemistry

    Background:

    • MicroRNA (miRNA) analysis is crucial for diagnostics.
    • Existing biosensor methods face limitations in sensitivity and specificity.
    • Two-dimensional nanoparticles (2D-nps) offer unique properties for biosensing applications.

    Purpose of the Study:

    • To develop a novel combinatorial nanosensor array for sensitive and specific miRNA detection.
    • To demonstrate the array's capability for simultaneous identification and quantification of multiple miRNA targets.
    • To validate the nanosensor's performance in complex biological matrices.

    Main Methods:

    • Assembly of a nanosensor array using three types of 2D-nps (WS2, MoS2, nGO) and five fluorescently labeled ssDNA probes.
    • Utilizing noncovalent interactions and fluorescence recovery measurements for target-probe interactions.
    • Employing partial least squares discriminant analysis (PLSDA) for data processing and target discrimination.

    Main Results:

    • Simultaneous discrimination of nine miRNA analogues with single nucleotide differences within 2 hours.
    • Accurate identification and quantification of unknown targets at unknown concentrations.
    • Successful detection of targets in a complex human urine matrix.
    • Demonstrated discrimination of target mixtures with varying ratios.

    Conclusions:

    • The developed combinatorial nanosensor array provides a powerful tool for miRNA analysis.
    • This approach overcomes limitations of traditional bind-and-release models for 2D-np biosensors.
    • The nanotechnology enables broad detection opportunities for 2D-nps in complex biological samples.