Amplification-free dnazyme-based point-of-care DNA/RNA detection platform

Overview

Cromerix is developing an innovative point-of-care detection platform for DNA and RNA targets that eliminates the need for nucleic acid amplification. Current DNAzyme-based detection methods rely on amplification techniques that are reagent-intensive, costly, and operationally complex. Cromerix's approach addresses these limitations by integrating target recognition and signal amplification into a single probe within a single sample tube.

The technology centers on a patent-pending dual-function probe in which one segment binds specifically to the target sequence while the other segment is a DNAzyme that becomes activated upon binding. Once activated, the DNAzyme cleaves multiple fluorophore-labeled substrate molecules successively, progressively turning on fluorescence and amplifying signal intensity without any additional reagents or steps. This enables sensitive quantitative detection in a streamlined workflow suitable for point-of-care settings.

Technical specifications

Core innovation:

• Single-step detection combining target identification and fluorescence signal transduction in one tube
• Dual-function probe design with binding-triggered structural reconstruction that activates DNAzyme activity only upon specific target binding
• DNAzyme cleaves multiple substrate molecules successively, providing intrinsic signal amplification
• Substrate molecules contain a fluorophore and quencher, producing fluorescence turn-on upon cleavage

Performance targets:

• Estimated sensitivity of approximately 500 attomolar for DNA/RNA detection
• Target detection time within 5 minutes after optimization
• Amplification-free workflow, reducing reagent costs and complexity

Customization approach:

The identification segment of the dual probe is tailored to the partner's specific DNA/RNA target sequence, while the transduction segment (DNAzyme and fluorophore-quencher substrate) remains unchanged across applications. This modular design enables rapid adaptation to diverse diagnostic targets.

Technology readiness level

The probe design has been validated using in-silico tools and is supported by literature evidence for the underlying DNAzyme properties. Experimental validation is currently underway. Performance projections are based on a recently reported mathematical model for DNAzyme-based detection kinetics. Future validation activities include demonstrating fluorescence turn-on using a partner-provided target sequence, optimizing probe and substrate concentrations and buffer conditions for reaction kinetics, and conducting limit-of-detection studies. The technology is at an early-to-mid stage of development with a clear pathway toward point-of-care deployment.