Pushbutton-activated microfluidic cell-free biosensor for multiplexed pathogen detection

Area of Science:

• Biotechnology
• Microfluidics
• Biosensors

Background:

• Pathogen detection is crucial for public health.
• Existing methods often lack multiplexing capabilities or require complex instrumentation.
• Cell-free systems offer rapid and adaptable diagnostic solutions.

Purpose of the Study:

• To develop a novel cell-free biosensor for simultaneous detection of multiple 16S rRNAs.
• To integrate multi-step cell-free protein synthesis into a single microfluidic device.
• To create an accessible and intuitive diagnostic platform.

Main Methods:

• Development of a multiplexed pushbutton-activated microfluidic device (mPAMD).
• Integration of target-responsive cell-free protein synthesis with finger-pumping mechanism.
• Design and optimization of microchannels for efficient mixing and reagent distribution.
• Incorporation of multiplexed detection zones with pathogen-specific probes.

Main Results:

• Simultaneous detection of multiple 16S rRNAs achieved in a single device.
• Achieved a detection limit for 16S rRNA ranging from 1.69 to 7.39 pM.
• Demonstrated high sensitivity and specificity for pathogen identification.
• System corresponds to approximately 10^4 to 10^5 CFU mL^-1 of pathogens.

Conclusions:

• The mPAMD provides a simple, intuitive, and accessible multiplexed diagnostic platform.
• The developed cell-free biosensor addresses the demand for sensitive and specific pathogen detection.
• This technology holds potential for rapid point-of-care diagnostics.