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CRISPR/Cas12a-mediated fluorescent aptasensor based on DNA walker amplification for oxytetracycline detection
Hualin Guo1, Pengfei Ma2, Xiaoze Dong1
1School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.
Biosensors & Bioelectronics
|December 7, 2024
Summary
A novel aptasensing platform detects oxytetracycline (OTC) using CRISPR/Cas12a and a DNA walker. This method achieves sensitive fluorescence detection of OTC in food samples, addressing environmental and health concerns.
Area of Science:
- Analytical Chemistry
- Biotechnology
- Environmental Science
Background:
- Oxytetracycline (OTC) contamination in the environment and food poses health risks.
- Understanding OTC aptamer molecular dynamics is crucial for developing detection methods.
Purpose of the Study:
- To develop a sensitive and efficient aptasensing platform for oxytetracycline (OTC) detection.
- To address the low efficiency of CRISPR/Cas12a systems in biosensing applications.
Main Methods:
- Utilized DNA walker amplification to enhance CRISPR/Cas12a activity.
- Integrated nanoclusters (NCs) with covalent organic framework (COF) for a "turn-on" fluorescence detection.
- Studied molecular dynamics of OTC aptamers to identify key base fragments.
Main Results:
- Achieved a linear detection range of 0.05-50 μM for OTC.
- Established a low detection limit of 0.041 μM for OTC.
- Demonstrated successful quantitative detection of OTC in milk and honey with high recovery rates (95.67%-109.70%).
Conclusions:
- The developed aptasensing platform offers excellent analytical performance for OTC detection.
- The method shows significant practical application prospects for monitoring OTC in food products.
- The study provides theoretical guidance for designing advanced aptamer-based detection systems.
