Multiplexed food-borne pathogen detection using an argonaute-mediated digital sensor based on a...

Area of Science:

• Food safety
• Molecular diagnostics
• Microbiology

Background:

• Accurate, sensitive, and multiplexed detection of food-borne pathogens is essential for food safety.
• Current methods often require DNA amplification, adding complexity and time.
• There is a need for rapid, accurate, and field-deployable pathogen detection systems.

Purpose of the Study:

• To develop a DNA-amplification-free nucleic acid detection assay for multiplexed and ultrasensitive detection of food-borne pathogens.
• To utilize a digital carrier system (d-MAGIC) incorporating mesophilic Clostridium butyricum argonaute and magnetic beads.
• To enable simultaneous detection of multiple pathogens with AI-driven data interpretation.

Main Methods:

• Developed a digital DNA-amplification-free nucleic acid detection assay using mesophilic Clostridium butyricum argonaute and magnetic beads in a digital carrier system (d-MAGIC).
• Employed Clostridium butyricum argonaute's two-guide accurate cleavage activity for precise targeting and cleavage of fluorescence-quencher reporters.
• Utilized fluorescence-encoded magnetic beads as programmable multi-probes for simultaneous pathogen detection.

Main Results:

• Achieved multiplexed and ultrasensitive detection of three food-borne pathogens without DNA amplification.
• Demonstrated a wide detection range from 10^1 to 10^7 CFU/ml.
• Obtained a low limit of detection of 6 CFU/ml for food-borne pathogens.

Conclusions:

• The d-MAGIC system offers a next-generation strategy for accurate and convenient pathogen detection.
• This DNA-amplification-free approach simplifies the detection process and enhances sensitivity.
• The developed assay has significant potential for improving food safety risk assessment.