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Double-probe signal enhancing strategy for toxin aptasensing based on rolling circle amplification
Ping Tong1, Wei-Wei Zhao, Lan Zhang
1The State Key Lab of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210093, China.
Biosensors & Bioelectronics
|January 25, 2012
Summary
A new electrochemical method uses aptamer-based rolling circle amplification (RCA) and magnetic beads (MBs) to detect Ochratoxin A (OTA) with high sensitivity. This innovation offers a promising tool for identifying OTA in food products like red wine.
Area of Science:
- Biosensing
- Analytical Chemistry
- Molecular Biology
Background:
- Ochratoxin A (OTA) is a prevalent mycotoxin found in food, posing significant health risks.
- Accurate and sensitive detection methods for OTA are crucial for food safety and quality control.
Purpose of the Study:
- To develop a highly sensitive electrochemical method for Ochratoxin A (OTA) determination.
- To leverage aptamer-based rolling circle amplification (RCA) and magnetic beads (MBs) for enhanced detection.
Main Methods:
- Immobilization of capture DNA onto magnetic beads (MBs) for OTA aptamer and padlock DNA hybridization.
- OTA-mediated DNA circularization and T4 ligase-catalyzed template formation.
- Linear rolling circle amplification (RCA) using phi29 DNA polymerase to amplify the signal.
- Detection via electrochemical readout using quantum dots (QDs) labeled DNA probes.
Main Results:
- The developed method achieved a sensitive detection of OTA down to 0.2 pg mL(-1).
- The assay demonstrated a wide dynamic range spanning over 4 orders of magnitude.
- Successful determination of OTA in red wine samples, indicating practical applicability.
Conclusions:
- The novel strategy combining OTA-mediated DNA circularization, RCA, and double signal probes offers a sensitive and reliable method for OTA detection.
- This electrochemical approach shows significant potential for routine analysis of OTA in real food samples.
