A multi-valve centrifugal microfluidic for Mycoplasma pneumoniae detection

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.