A probe-mediated fluorescent biosensor for MC-LR detection using exonuclease III as a signal amplifier
Yuyan Wang1, Ying Zeng1, Xiaoya Ren1
1Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, Hengyang Medical School, University of South China, Hengyang 421001, China. yangfeilong@126.com.
Abstract:
Microcystin-lr (MC-LR) is one of the most toxic and ubiquitous microcystins (MCs) released by cyanobacteria. Exposure to MC-LR can cause multiple organ damage and even death of the organism. Therefore, creating highly sensitive and dependable methods for detecting trace MC-LR is crucial. Herein, we developed a novel fluorescence aptasensor aided by exonuclease III (Exo III) for the highly sensitive detection of MC-LR. In the presence of MC-LR, the affinity interaction between MC-LR and aptamer A was triggered, leading to the release of blocker B. This unbound blocker can initiate Exo III-mediated signal amplification to digest the probe H, thereby recovering the fluorescence signal for readout. The proposed Exo III-assisted sensing platform demonstrated remarkable sensitivity and selectivity, achieving a limit of detection (LOD) of 0.37 ng L-1. Furthermore, it is robust and has been effectively utilized on water samples, achieving acceptable recovery rates (95.04-107.01%). With excellent sensitivity, high selectivity, efficient signal amplification, and fluorescence readout, the proposed biosensor offered a new and reliable alternative for the detection of trace MC-LR in the environment and the early warning of algal toxins.
Insights
A new fluorescence biosensor accurately detects trace amounts of microcystin-LR (MC-LR), a potent cyanotoxin. This novel method, utilizing exonuclease III (Exo III) for signal amplification, offers a reliable tool for environmental monitoring and early warning of algal blooms.
Area of Science:
- Environmental Science
- Biotechnology
- Analytical Chemistry
Background:
- Microcystin-LR (MC-LR) is a highly toxic cyanotoxin causing significant health risks.
- Sensitive and reliable detection methods for MC-LR are essential for public health and environmental safety.
Purpose of the Study:
- To develop a novel, highly sensitive fluorescence aptasensor for detecting trace levels of MC-LR.
- To leverage exonuclease III (Exo III) for enhanced signal amplification in MC-LR detection.
Main Methods:
- A fluorescence aptasensor was designed utilizing MC-LR specific aptamers and a blocker molecule.
- Exonuclease III (Exo III) was employed for signal amplification through probe digestion upon MC-LR binding.
- The biosensor's performance was evaluated for sensitivity, selectivity, and application in real water samples.
Main Results:
- The developed aptasensor achieved a low limit of detection (LOD) of 0.37 ng L⁻¹ for MC-LR.
- The biosensor demonstrated high sensitivity and selectivity for MC-LR detection.
- Successful application in water samples with recovery rates ranging from 95.04% to 107.01%.
Conclusions:
- The Exo III-assisted fluorescence aptasensor provides a sensitive, selective, and reliable method for detecting trace MC-LR.
- This biosensor offers a promising alternative for environmental monitoring and early warning systems for harmful algal blooms.
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