Aptamer-Based Dual-Cascade Signal Amplification System Lights up G-Quadruplex Dimers for Ultrasensitive Detection of Domoic Acid

TL;DR

This paper introduces a highly sensitive fluorescent sensor for detecting domoic acid in shellfish using a dual-signal amplification system.

Contribution

The novel dual-cascade signal amplification system combines G-quadruplex dimers and carbon nanotubes for ultrasensitive detection.

Findings

The sensor detects domoic acid with a detection limit of 1.1 ng/mL.

Detection occurs within 5 minutes, making it rapid and practical for real-world applications.

The system uses a hairpin-structured aptamer and thioflavin T for signal amplification.

Abstract

In recent years, harmful algal blooms have led to frequent occurrences of shellfish toxin contamination, posing a significant threat to the safety of aquatic products and public health. As a potent neurotoxin, domoic acid (DA) can accumulate in shellfish, highlighting the urgent need for rapid and highly sensitive detection methods. In this study, we developed a fluorescent aptasensor based on a dual-signal amplification system by combining G-quadruplex (G4) dimers with multi-walled carbon nanotubes (CNTs). The sensor is designed with a hairpin-structured aptamer as the recognition probe, where short multi-walled CNTs serve as both a fluorescence quencher and platform, and G4 dimers are incorporated into the sensing interface to enhance signal output. In the absence of the target, the hairpin-structured aptamer remains closed, keeping the fluorescence signal “off”. Upon binding to DA, the aptamer undergoes a specific conformational change that exposes the G4-dimer sequence. The exposed sequence then binds to thioflavin T (ThT), which in turn generates a greatly enhanced fluorescence signal, leading to a substantial fluorescence enhancement and completing the second stage of the cascade amplification. Under optimal conditions, the constructed sensor achieves rapid detection of DA within 5 min, with a low detection limit of 1.1 ng/mL. This work presents a valuable tool for the rapid and sensitive detection of DA in shellfish, with promising applications in marine environmental monitoring and food safety regulation.

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