Triplex DNA-based aggregation-induced emission probe: A new platform for hybridization chain reaction-based...

Area of Science:

• Biotechnology
• Molecular Biology
• Analytical Chemistry

Background:

• Hybridization chain reaction (HCR) offers sensitive nucleic acid amplification but faces challenges with fluorophore blocking and aggregation-caused quenching (ACQ).
• Traditional fluorescence sensing methods using HCR are limited by these inherent issues, impacting signal output and sensitivity.
• Development of novel probes is crucial to overcome existing limitations in HCR-based fluorescence assays.

Purpose of the Study:

• To design and develop a novel triplex DNA-based aggregation-induced emission probe (AIE-P) for enhanced HCR fluorescence sensing.
• To address the limitations of traditional fluorophores and the blocking effect in HCR assays.
• To establish a new platform for sensitive and specific detection using HCR technology.

Main Methods:

• Synthesized an AIE-P by conjugating AIE fluorophores to oligonucleotide termini via an amido bond.
• Utilized the AIE-P to capture HCR products, forming triplex DNA structures that enhance fluorescence emission.
• Developed a turn-on fluorescence assay for detecting Vibrio parahaemolyticus.

Main Results:

• The AIE-P facilitated a turn-on fluorescence response with a high signal-to-noise ratio and excellent amplification.
• Achieved sensitive detection of Vibrio parahaemolyticus within a concentration range of 10^2-10^6 CFU mL^-1.
• Established a low limit of detection of 39 CFU mL^-1, demonstrating high sensitivity and specificity.

Conclusions:

• The triplex DNA-based AIE probe offers a universal molecular tool for advancing HCR-based fluorescence sensing methods.
• This approach overcomes key limitations of existing HCR fluorescence assays, improving sensitivity and signal efficiency.
• The developed assay demonstrates practical applicability and high specificity for bacterial detection.