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A Relay CRISPR/Cas12a System for Extraction-Free and Ultrasensitive Detection of Staphylococcus aureus in Complex
Kun Wang1, Wanli Wang1, Yufeng Hu1
1Xiangya School of Public Health, Central South University, Changsha 410013, Hunan, China.
Analytical Chemistry
|January 7, 2026
Summary
This study presents a novel biosensor for detecting Staphylococcus aureus (SA) in food. The system uses a DNA walker and CRISPR/Cas12a for ultrasensitive bacterial identification, enabling both lab and on-site analysis.
Area of Science:
- Biotechnology
- Food Safety
- Molecular Diagnostics
Background:
- Staphylococcus aureus (SA) is a major foodborne pathogen found in nutrient-rich foods.
- Detecting SA rapidly and sensitively in complex food samples is challenging.
Purpose of the Study:
- To develop a relay biosensing system for ultrasensitive detection of SA-16S rDNA.
- To enable both laboratory and on-site detection of SA in food matrices.
Main Methods:
- A magnetic bead-anchored APE1 DNA walker was combined with an asymmetric CRISPR/Cas12a system.
- The system utilized spatially decoupled reactions with differential crRNAs for cascade amplification.
- Dual-mode fluorescence and electrochemical readouts were employed.
Main Results:
- The biosensor detected SA-16S rDNA at 7.5 aM (fluorescence) and 1.0 fM (electrochemistry).
- It detected SA at 4 CFU/mL (fluorescence) and 34 CFU/mL (electrochemistry).
- The system demonstrated efficient separation of SA targets from complex food matrices.
Conclusions:
- The developed biosensing strategy is modular and programmable for bacterial identification.
- This facilitates advanced nucleic acid analysis in complex biological samples.
- It offers a promising approach for food safety applications.
