Glucosemeter-based aptasensor with an efficient and green system driven by CRISPR/Cas12a for ZEN detection.

Area of Science:

• Biosensing
• Molecular Diagnostics
• Analytical Chemistry

Background:

• Zearalenone (ZEN) is a mycotoxin posing risks to food safety and human health.
• Existing ZEN detection methods often lack cost-effectiveness and accessibility.
• There is a need for rapid, sensitive, and field-deployable ZEN monitoring tools.

Purpose of the Study:

• To develop a novel, cost-effective, and readily available aptasensor platform for zearalenone (ZEN) detection.
• To utilize the clustered regularly interspaced short palindromic repeats/associated protein 12a (CRISPR/Cas12a) system for enhanced detection sensitivity.
• To integrate a glucosemeter for simplified and accessible quantification of ZEN levels.

Main Methods:

• A magnetic bead (MB)-based aptasensor was constructed using MB@Apt-cDNA and MB@invertase probes.
• The system leverages CRISPR/Cas12a's DNA-cleaving activity triggered by ZEN.
• Generated glucose signals from sucrose conversion by released invertase were measured using a glucosemeter.

Main Results:

• The aptasensor achieved a limit of detection (LOD) of 0.218 ng/mL for ZEN.
• The detection range spanned from 0.218 to 109.89 ng/mL with high specificity (cross-reactivity < 6.81%).
• The platform demonstrated high accuracy and reliability in real agricultural byproduct samples (AUC = 0.979).

Conclusions:

• The proposed CRISPR/Cas12a and glucosemeter-assisted aptasensor offers an efficient and accessible method for ZEN detection.
• This platform provides an alternative pathway for improving detection performance in resource-limited settings.
• The study suggests a versatile strategy for applying CRISPR/Cas12a and glucosemeter detection to other environmental pollutants.