CRISPR/Cas12a-based electrochemical biosensor for highly sensitive detection of cTnI

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Cardiovascular Research

Background:

• Myocardial injury and cardiovascular diseases (CVDs) require rapid and accurate diagnostic tools.
• Current detection methods for cardiac troponin I (cTnI) may lack the sensitivity or speed needed for instant diagnosis.

Purpose of the Study:

• To develop a highly sensitive and specific electrochemical biosensor for the rapid detection of cTnI.
• To leverage the CRISPR/Cas12a system for enhanced diagnostic capabilities in cardiovascular disease detection.

Main Methods:

• Fabrication of an electrochemical biosensor using magnetic nanoparticles, aptamers, and the CRISPR/Cas12a/crRNA system.
• Electrochemical detection of cTnI based on aptamer binding, subsequent release of a DNA probe, and CRISPR/Cas12a-mediated trans-cleavage.
• Utilizing methylene blue-modified DNA as an electrochemical signal indicator.

Main Results:

• The biosensor demonstrated high specificity and sensitivity for cTnI detection.
• Achieved a low detection limit of 10 pg/mL for cTnI within a linear range of 100-50000 pg/mL.
• Successfully validated the platform's performance in detecting cTnI in human serum samples.

Conclusions:

• The developed CRISPR/Cas12a-based electrochemical biosensor is a valuable tool for the instant diagnosis, prognosis, and treatment of CVDs.
• The synergistic effects of aptamer recognition, CRISPR/Cas12a activity, and magnetic nanoparticle separation contribute to the biosensor's efficacy.
• This platform offers a promising approach for point-of-care diagnostics in cardiovascular medicine.