Facile and Ultrafast Microfluidic Photothermal PCR for Autonomous and Quantitative Point-of-Care Pathogen Detection

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Point-of-Care Testing

Background:

• Accurate and rapid pathogen detection is crucial for infectious disease diagnosis, food safety, and environmental monitoring.
• Existing point-of-care (POC) molecular diagnostic systems often require complex fabrication and lengthy processing times.
• There is a need for simplified, autonomous systems for on-site and in-home pathogen detection.

Purpose of the Study:

• To develop a simplified, automated microfluidic photothermal quantitative polymerase chain reaction (AMP-qPCR) system.
• To enable rapid, autonomous sample processing including lysis, purification, and real-time PCR.
• To demonstrate the system's efficiency and accuracy for bacterial quantification and clinical pathogen detection.

Main Methods:

• Development of a magnetically-driven, multi-chamber microfluidic cartridge.
• Integration of a low-cost black tape as a photothermal-responsive engine for rapid heating.
• Real-time fluorescence quantitative polymerase chain reaction (qPCR) for pathogen detection.
• Validation using bacterial DNA extraction and quantification, and detection of *Neisseria gonorrhoeae* in clinical samples.

Main Results:

• The AMP-qPCR system achieved high-efficiency bacterial DNA extraction and quantitative PCR in 18.5 minutes.
• Accurate quantification of bacteria concentrations ranging from 10^8 to 10^2 CFU·mL^-1 was demonstrated.
• The system showed matched performance to benchtop automated machines in detecting *Neisseria gonorrhoeae* from clinical specimens.

Conclusions:

• The developed AMP-qPCR system offers a simplified, autonomous, and rapid solution for POC pathogen detection.
• This technology enhances the accessibility of molecular diagnostics for on-site and in-home testing.
• The platform holds significant potential for improving infectious disease diagnosis and public health surveillance.