100 Section - Development of a point-of-care testing for Stroke-Related Nucleic Acid Markers Based on Cas Protein Precision Regulation System

Stroke is the neurological disease with the highest disability rate in China. The recovery process varies significantly between individuals, making existing assessment methods such as imaging and functional scales ineffective in dynamically and sensitively reflecting neurological recovery. However, traditional detection methods rely on amplification and reverse transcription, resulting in limited sensitivity and complex procedures, making them difficult to apply to point-of-care testing.

This project aims to develop a nucleic acid detection system with intelligent feedback control capabilities based on the precise regulation of Cas proteins, enabling highly sensitive, low-background, and amplification-free detection of stroke-related miRNAs.

Design:

The research will construct a CRISPR molecular detection platform. Ultimately, the project aims to develop a portable point-of-care computerized computer (POCT) platform with clinical potential for monitoring rehabilitation outcomes and evaluating personalized interventions. This project will provide a new, efficient, economical and translatable molecular detection method for precise stroke rehabilitation, and promote the clinical transformation of Cas protein molecular diagnostic technology in the field of neurorehabilitation.

Results: We successfully developed a CRISPR/Cas system for detecting miRNAs. This system successfully detected miR-220b, miR-140-3p, and miR-429 in vitro. It is expected that this system could be used for in vitro diagnostics of stroke-related miRNAs.

Conclusion:

In summary, this project successfully established a CRISPR/Cas-based nucleic acid detection platform for the identification of stroke-related miRNAs. The system achieved reliable detection of key biomarkers such as miR-220b, miR-140-3p, and miR-429 in vitro. These results support the feasibility of integrating this detection module into a portable point-of-care testing (POCT) device for stroke-related nucleic acid markers.