FINDER converts zero-background kinetic fingerprinting into area-scalable attomolar biomarker detection

Area of Science:

• Biomolecular analysis
• Single-molecule detection
• Analytical chemistry

Background:

• Analytical sensitivity is often limited by background noise, hindering accurate detection.
• Existing methods struggle to balance sensor area with false positive accumulation.
• Developing sensitive, amplification-free detection methods is crucial for early disease diagnosis.

Purpose of the Study:

• Introduce FINDER (Fluorogenic INstantaneous Digital Enumeration and Recognition), a novel single-molecule platform.
• Enable rapid molecular classification and quantification under near-zero-background conditions.
• Achieve amplification-free detection with high sensitivity and scalability.

Main Methods:

• Utilizes kinetic fingerprinting and fluorogenic transient probes for molecular classification.
• Suppresses solution and surface-associated background noise.
• Employs multidimensional kinetic filtering for discrimination of variants.

Main Results:

• Achieved attomolar detection limits for miRNA cancer biomarker hsa-miR-16.
• Demonstrated generalization to HPV16 DNA detection and RNA/DNA co-profiling.
• Enabled rapid discrimination of EGFR single-nucleotide variants.
• Showcased area-scalable sensitivity for amplification-free quantification.

Conclusions:

• FINDER offers a powerful approach for sensitive, rapid, and amplification-free biomarker quantification.
• The platform enables semi-automated attomolar biomarker counting in practical workflows.
• FINDER's kinetic specificity translates to area-scalable sensitivity, advancing molecular diagnostics.