Digital Microfluidic qPCR Cartridge for SARS-CoV-2 Detection

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Microfluidics

Background:

• Point-of-care (POC) testing is crucial for managing pandemics like COVID-19, aiding in health monitoring, transmission reduction, and contact tracing.
• Digital microfluidics (DMF) offers a platform for developing compact and efficient diagnostic devices.

Purpose of the Study:

• To develop a mass-producible, disposable POC cartridge for SARS-CoV-2 N gene detection using DMF-based real-time quantitative polymerase chain reaction (qPCR).
• To optimize critical performance parameters of the DMF cartridge for accurate and efficient molecular diagnostics.

Main Methods:

• Development of a disposable DMF cartridge designed for high-throughput sample processing.
• Optimization of droplet volume consistency, temperature uniformity, and fluorescence intensity linearity on the DMF platform.
• Utilizing two primer-probe sets (N1 and N2) for SARS-CoV-2 N gene detection via qPCR on the cartridge.

Main Results:

• The DMF cartridge demonstrated high accuracy and efficiency in detecting the SARS-CoV-2 N gene.
• Optimized parameters led to improved droplet control, thermal stability, and signal linearity.
• The cartridge design supports multiplexed testing and internal controls due to multiple droplet tracks.

Conclusions:

• The developed DMF-based POC cartridge is a viable tool for accurate and efficient SARS-CoV-2 detection.
• The cartridge's design facilitates mass production and multiplexed testing capabilities, suitable for pandemic response.
• This technology advances POC diagnostics for infectious diseases, supporting public health initiatives.