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Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction
Published on: October 6, 2022
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A novel biosensor based on antibody controlled isothermal strand displacement amplification (ACISDA) system
Xiao Liu1, Han Yang2, Zehua Xu1
1Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Tianjin Institute of Environmental and Operational Medicine, Tianjin, 300050, PR China.
Biosensors & Bioelectronics
|April 16, 2022
Summary
Antibiotic overuse necessitates rapid detection methods. A new antibody-controlled isothermal chain displacement amplification (ACISDA) system offers sensitive detection of antibiotic residues, ensuring food and environmental safety.
Area of Science:
- Analytical Chemistry
- Biotechnology
- Environmental Science
Background:
- Antibiotic overuse poses risks to human health via food and environmental residues.
- Sensitive and rapid detection methods for antibiotics are crucial.
Purpose of the Study:
- To develop a novel, highly sensitive, and rapid detection strategy for small molecules like antibiotics.
- To establish an antibody-controlled isothermal chain displacement amplification (ACISDA) system for antibiotic detection.
Main Methods:
- Utilized antibody-controlled isothermal chain displacement amplification (ACISDA) inspired by allosteric transcription factors.
- Integrated nicking endonuclease, Klenow Fragment polymerase, specific antibodies, and antigen-labeled DNA to synthesize G-quadruplex.
- Employed thioflavine T (ThT) for fluorescent signal generation upon G-quadruplex formation.
Main Results:
- Developed an ACISDA system for norfloxacin detection using anti-norfloxacin antibody and norfloxacin-conjugated DNA.
- Achieved a linear detection range of 0.1–500 ng/mL for norfloxacin with a low detection limit of 0.04 ng/mL.
- Demonstrated good performance in detecting norfloxacin in real samples, highlighting the system's applicability.
Conclusions:
- The ACISDA system provides a sensitive, rapid, and versatile platform for small molecule detection.
- The system is washing-free, immobilization-free, and ready-to-use, adaptable for various small molecules by modifying DNA and antibody components.
