A Portable Dual-Mode Microfluidic Device Integrating RT-qPCR and RT-LAMP for Rapid Nucleic Acid Detection in...

Point-of-care testing (POCT) has emerged as a vital diagnostic approach in emergency medicine, primary care, and resource-limited environments because of its convenience, affordability, and capacity to provide immediate results. Here, we present a multifunctional portable nucleic acid detection platform integrating reverse transcription polymerase chain reaction (RT-qPCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP) within a unified microfluidic device. The system leverages Tesla-valve-based passive flow control to enhance reaction efficiency and operational simplicity. A four-channel optical detection unit allows for multiplex fluorescence quantification (CY5, FAM, VIC, ROX) and has high sensitivity and reproducibility for RT-LAMP. The compact design reduces the overall size by approximately 90% compared with conventional qPCR instruments. For RT-PCR, the system achieves a detection limit of 2.0 copies μL^-1 and improves analytical efficiency by 27%. For RT-LAMP, the detection limit reaches 2.95 copies μL^-1 with a 14% enhancement in analytical efficiency. Compared with commercial qPCR instruments, the device maintains equivalent quantitative accuracy despite significant miniaturization, ensuring reliable performance in decentralized testing. Furthermore, the total RT-LAMP assay time is reduced from more than two hours to 42 min, enabling truly rapid molecular diagnostics. This dual-mode platform offers a flexible, scalable strategy for bridging laboratory-grade molecular assays with real-time POCT applications, supporting early disease detection and epidemic surveillance.