A Digital Microfluidic RT-qPCR Platform for Multiple Detections of Respiratory Pathogens

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Infectious Disease Research

Background:

• The COVID-19 pandemic highlights the critical need for rapid, accurate diagnostics to control infectious disease spread.
• Traditional nucleic acid detection methods like RT-PCR require specialized labs and personnel, limiting accessibility in remote areas.
• Digital microfluidic (DMF) technology offers a promising alternative for multiplexed pathogen detection with reduced sample requirements and simplified operation.

Purpose of the Study:

• To develop and validate a novel digital microfluidic (DMF) based RT-qPCR platform for the simultaneous detection of multiple respiratory pathogens.
• To assess the diagnostic performance of the DMF platform against established off-chip RT-PCR methods.

Main Methods:

• Development of a novel digital microfluidic (DMF) platform integrated with RT-qPCR for multiplexed nucleic acid detection.
• Simultaneous detection of eleven key respiratory pathogens, including SARS-CoV-2, influenza viruses, and other common respiratory agents.
• Evaluation of diagnostic performance using positive plasmid samples and clinical specimens, comparing results with individual off-chip RT-PCR.

Main Results:

• The DMF RT-qPCR platform demonstrated high diagnostic accuracy, with a true positive rate of 93.33% and true negative rate of 100%.
• The system achieved excellent performance metrics, including 100% positive predictive value, 99.56% negative predictive value, and 99.85% overall accuracy.
• The limit of detection ranged from 12 to 150 copies per test, indicating high sensitivity for pathogen identification.

Conclusions:

• The developed on-chip DMF RT-qPCR system is a cost-effective, highly sensitive, and specific solution for multiplex respiratory pathogen detection.
• This technology significantly enhances the potential for timely and effective clinical management of respiratory infections, particularly in resource-limited settings.
• The DMF platform offers a user-friendly and efficient alternative to conventional methods, improving accessibility to advanced diagnostics.