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Related Experiment Video

Updated: Oct 2, 2025

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Published on: March 31, 2021

Digital Microfluidic qPCR Cartridge for SARS-CoV-2 Detection.

Kuan-Lun Ho¹, Hong-Yu Liao², Helene Minyi Liu³

¹Department of Mechanical and Nuclear Engineering, Kansas State University, Manhattan, KS 66506, USA.

|February 25, 2022

Summary

A new disposable point-of-care (POC) cartridge uses digital microfluidics (DMF) and real-time quantitative polymerase chain reaction (qPCR) to accurately detect the SARS-CoV-2 N gene for COVID-19 testing.

Keywords:

COVID-19 SARS-CoV-2 digital microfluidics droplet qPCR electrowetting

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Last Updated: Oct 2, 2025

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

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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.