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"Aptamer-locker" DNA coupling with CRISPR/Cas12a-guided biosensing for high-efficiency melamine analysis
Bin Qiao1, Jiakun Xu2, Wenhao Yin3
1Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University, China.
Biosensors & Bioelectronics
|April 13, 2021
Summary
This study introduces a novel DNA sensor using aptamer-lockers and CRISPR/Cas12a technology for fast and sensitive melamine detection. The method achieves a low limit of detection, crucial for ensuring food safety in products like infant milk.
Area of Science:
- Biotechnology and Biosensing
- Molecular Diagnostics
- Food Safety Analysis
Background:
- Melamine contamination poses significant risks to food safety, particularly in infant milk products.
- Existing detection methods for melamine can be time-consuming or lack the required sensitivity.
- CRISPR/Cas12a systems offer a powerful platform for highly specific molecular detection.
Purpose of the Study:
- To develop a sensitive and rapid biosensing method for melamine detection.
- To integrate aptamer-locker DNA with CRISPR/Cas12a for enhanced melamine analysis.
- To validate the method's performance in real-world food samples, such as milk.
Main Methods:
- Design and characterization of aptamer-locker DNA sensors using circular dichroism (CD) and isothermal titration calorimetry (ITC).
- Optimization of detection parameters for CRISPR/Cas12a-based biosensing.
- Application of the developed sensor for melamine detection in whole milk samples using a portable fluorimeter.
Main Results:
- Achieved a low limit of detection (LOD) of 38 nM for melamine, below regulatory thresholds.
- Demonstrated high selectivity for melamine over structurally similar compounds like cyanuric acid.
- Enabled rapid (under 20 minutes) and on-site melamine analysis in milk, with or without pretreatment.
Conclusions:
- The combined aptamer-locker and CRISPR/Cas12a approach provides a sensitive, rapid, and selective method for melamine detection.
- This biosensing strategy is suitable for on-site food safety monitoring, especially for milk products.
- The platform is adaptable for detecting other non-nucleic acid targets by modifying the aptamer component.
