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Electrowetting-based Digital Microfluidics Platform for Automated Enzyme-linked Immunosorbent Assay
Published on: February 23, 2020
A magnet-actuated microfluidic array chip for high-throughput pretreatment and amplification and detection of
Mengfan Zhou1, Huiying Su2,1, Bangfeng Wang1
1The Key Laboratory for Biomedical Photonics of MOE at Wuhan National Laboratory for Optoelectronics - Hubei Bioinformatics & Molecular Imaging Key Laboratory, Systems Biology Theme, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China. xfeng@mail.hust.edu.cn.
Abstract:
The outbreak of global infectious diseases has posed a significant threat to public health, requiring the rapid and accurate diagnosis of pathogens promptly for the society to implement immediate control measures to prevent widespread pandemics. In this work, a magnet-actuated microfluidic array chip (MMAC) is developed with integrated sample processing and nucleic acid amplification for the rapid detection of multiple pathogens by loop-mediated isothermal amplification. In comparison to previous works, where fluid control was dependent on external equipment or finger-based manual pressing, the fluid control of the MMAC is realized by magnetically actuating a ferric oxide (Fe3O4) doped polydimethylsiloxane (PDMS) layer that separates the sample from the LAMP reagent in a high-throughput manner, which not only reduces the complexity of fluid control but also enhances the repeatability of detection by eliminating variations in operation by different users. Examination with a testing sample containing Salmonella typhimurium and Escherichia coli showed high specificity for pathogen detection without cross-contamination. The lowest detection concentration was 5.2 copies per μL for Salmonella typhimurium with a detection time of 60 min. The proposed method demonstrated the simultaneous detection of multiple pathogens, which is potentially helpful in applications of immediate diagnosis.
Insights
This study introduces a novel magnet-actuated microfluidic array chip for rapid, simultaneous detection of multiple pathogens using loop-mediated isothermal amplification. The device offers enhanced control and repeatability for timely infectious disease diagnosis.
Area of Science:
- Biotechnology
- Microfluidics
- Molecular Diagnostics
Background:
- Global infectious disease outbreaks necessitate rapid pathogen identification for effective pandemic control.
- Current diagnostic methods often lack speed, high-throughput capability, or ease of use.
Purpose of the Study:
- To develop a magnet-actuated microfluidic array chip (MMAC) for integrated sample processing and nucleic acid amplification.
- To enable rapid, high-throughput, and repeatable detection of multiple pathogens.
Main Methods:
- Development of an MMAC utilizing magnetically actuated ferric oxide-doped polydimethylsiloxane for fluid control.
- Integration of sample processing and loop-mediated isothermal amplification (LAMP).
- Testing with Salmonella typhimurium and Escherichia coli for specificity and sensitivity.
Main Results:
- The MMAC demonstrated high specificity, detecting pathogens without cross-contamination.
- Achieved a lowest detection concentration of 5.2 copies/μL for Salmonella typhimurium within 60 minutes.
- Successfully enabled simultaneous detection of multiple pathogens.
Conclusions:
- The MMAC offers a simplified, repeatable, and high-throughput solution for pathogen detection.
- This technology has potential for immediate and simultaneous diagnosis of infectious diseases.
- The magnetic actuation system overcomes limitations of external equipment or manual operation in microfluidic devices.
