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A multi-valve centrifugal microfluidic for Mycoplasma pneumoniae detection
Guangyao Chen1, Yu Lu1, Hao Shen2
1School of Mechanical and Electrical Engineering, Jiangsu Provincial Key Laboratory of Advanced Robotics, Soochow University, Suzhou, 215123, China. chenliguo@suda.edu.cn.
Abstract:
Mycoplasma pneumoniae is a widely prevalent infectious agent, and its rapid and accurate detection using isothermal amplification technology is crucial for maintaining both personal and public health safety. In recent years, microfluidic technology has demonstrated advantages over traditional clinical methods by reducing reagent consumption and shortening detection time. However, a challenge remains in balancing the need to shorten detection time with ensuring the adequacy of the amplification reaction. Minimizing detection time while guaranteeing the accuracy of the reaction is a key challenge. Therefore, this work designs a centrifugal microfluidic chip that integrates sample lysis, nucleic acid extraction, and amplification processes. The sequential release of the reaction solution is achieved by matching the release characteristics of Euler force valves, pneumatic centrifugal valves, and Euler force-driven siphon valves. The design of the delayed-release structure enhances the concentration of extracted nucleic acids, while precise control of the rotational speed allows for more rational allocation of detection time. Ultimately, qualitative detection of Mycoplasma pneumoniae is completed within 10 minutes. Concurrently, the nucleic acid concentration extracted by the chip reached 74 ng μl-1, with a minimum detection limit of 103 copies per ml. The optimization method for centrifugal microfluidic chip design proposed in this manuscript contributes to the promotion of this technology in clinical medicine.
Insights
This study presents a novel centrifugal microfluidic chip for rapid detection of Mycoplasma pneumoniae. The integrated system achieves accurate results in under 10 minutes, enhancing public health diagnostics.
Area of Science:
- Biomedical Engineering
- Molecular Diagnostics
- Microfluidics
Background:
- Mycoplasma pneumoniae is a common pathogen requiring rapid detection.
- Microfluidic technology offers advantages in speed and reagent efficiency over traditional methods.
- Balancing detection speed with amplification accuracy is a key challenge in microfluidic diagnostics.
Purpose of the Study:
- To design and validate a centrifugal microfluidic chip for integrated sample lysis, nucleic acid extraction, and amplification of Mycoplasma pneumoniae.
- To optimize the chip design for rapid detection while ensuring amplification efficiency.
- To demonstrate the clinical potential of the developed microfluidic system.
Main Methods:
- Development of a centrifugal microfluidic chip with integrated sample processing steps.
- Utilized Euler force valves, pneumatic centrifugal valves, and siphon valves for sequential reagent release.
- Incorporated a delayed-release structure to enhance nucleic acid concentration.
- Controlled rotational speed for optimized reaction time allocation.
Main Results:
- Qualitative detection of Mycoplasma pneumoniae was achieved in under 10 minutes.
- The chip extracted nucleic acids with a concentration of 74 ng/μl.
- A minimum detection limit of 10^3 copies/ml was established.
- The system demonstrated efficient integration of lysis, extraction, and amplification.
Conclusions:
- The developed centrifugal microfluidic chip enables rapid and accurate detection of Mycoplasma pneumoniae.
- The chip design optimizes nucleic acid concentration and detection time, addressing key challenges in microfluidic diagnostics.
- This technology shows promise for widespread clinical application in infectious disease diagnostics.
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