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Visualization of Surface-tethered Large DNA Molecules with a Fluorescent Protein DNA Binding Peptide
Published on: June 23, 2016
Binding-Induced DNA Dissociation Assay for Small Molecules: Sensing Aflatoxin B1
Lin Xu1,2, Hongquan Zhang2, Xiaowen Yan2
1Key Laboratory of Biology and Genetic Improvement of Oil Crops, Key Laboratory of Detection for Mycotoxins, Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, and National Reference Laboratory for Biotoxin Test , Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences , Wuhan 430062 , P. R. China.
A new sensor detects aflatoxin B1 (AFB1) using fluorescence. This method enhances signal-to-background ratio by 390% and achieves a low detection limit for AFB1, improving mycotoxin analysis.
Area of Science:
- Analytical Chemistry
- Biotechnology
- Food Safety
Background:
- Aflatoxin B1 (AFB1) is a potent mycotoxin requiring sensitive detection methods.
- Current detection methods may lack sensitivity or require complex procedures.
Purpose of the Study:
- To develop a novel fluorescence turn-on sensor for homogeneous detection of AFB1.
- To improve the signal-to-background ratio for enhanced sensitivity.
Main Methods:
- Utilized gold nanoparticles (AuNPs) functionalized with antibodies and assistant oligonucleotides (AO).
- Employed AFB1-bovine serum albumin (BSA) conjugates with fluorophore-labeled signal oligonucleotides (SO).
- Leveraged binding-induced intramolecular hybridization for fluorescence quenching and competitive release for signal-on detection.
Main Results:
- Achieved a 390% improvement in signal-to-background ratio due to enhanced fluorescence quenching.
- Established a detection limit of 2.3 nM for AFB1, meeting regulatory requirements.
- Demonstrated high recovery rates (86-102%) in spiked rice samples.
Conclusions:
- The developed sensor provides a sensitive and efficient method for AFB1 detection.
- The assay's design is adaptable for detecting other small molecules by changing antibody probes.
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