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Development of an Electrochemical DNA Biosensor to Detect a Foodborne Pathogen
Published on: June 3, 2018
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Label-Free Fluorescence Biosensor Constructed Based on Pyrococcus furiosus Argonaute System and Programmable DNAzyme
Xiru Zhang1, Manyan Qiu1, Jingwen Zhang1
1Key Laboratory of Dairy Science, Ministry of Education, Department of Food Science, Northeast Agricultural University, Harbin 150030, China.
Analytical Chemistry
|October 17, 2025
Summary
A novel biosensor using Pyrococcus furiosus Argonaute (PfAgo) and Aurora DNAzyme detects Salmonella typhimurium. This highly sensitive method offers rapid and reliable foodborne pathogen detection, crucial for public health and safety.
Area of Science:
- Biotechnology
- Biosensing
- Food Safety
Background:
- Salmonella typhimurium (S. typhimurium) is a major foodborne pathogen threatening human health and food safety.
- Sensitive and reliable detection methods for S. typhimurium are essential for preventing outbreaks and ensuring food security.
- Argonaute proteins offer high specificity and programmability for detecting pathogenic bacteria.
Purpose of the Study:
- To develop a novel, label-free fluorescent biosensing platform for sensitive S. typhimurium detection.
- To integrate Pyrococcus furiosus Argonaute (PfAgo) with a programmable Aurora DNAzyme for enhanced detection capabilities.
- To establish a new method for detecting pathogenic bacteria using Argonaute proteins and DNAzymes.
Main Methods:
- Designed a label-free signaling probe (engineered-Aurora, E-Aurora) by incorporating the target sequence into the Aurora DNAzyme's catalytic core.
- Utilized the E-Aurora probe's ability to catalyze 4-methylumbelliferyl phosphate (4-MUP) into a fluorescent compound (4-MU).
- Employed PfAgo, activated by S. typhimurium, to cleave E-Aurora, thereby inhibiting fluorescence signal generation.
Main Results:
- The PfAgo-activated cleavage of E-Aurora led to an inverse relationship between S. typhimurium concentration and fluorescence signal.
- The biosensor demonstrated high sensitivity for S. typhimurium detection, achieving a limit of detection as low as 1 CFU/mL.
- The integrated system leveraged the specific recognition of PfAgo and the catalytic efficiency of E-Aurora for sensitive detection.
Conclusions:
- Developed a novel label-free fluorescent biosensor based on PfAgo and Aurora DNAzyme for sensitive S. typhimurium detection.
- The biosensor offers a promising tool for rapid, reliable, and highly sensitive detection of foodborne pathogens.
- This work provides a new strategy for designing Argonaute-based fluorescence biosensors for pathogenic bacteria detection.
