Xiangyu Jiang

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.

This study addresses the issues of poor stability and difficulty in recovery of free horseradish peroxidase (HRP) by developing a multi-level composite immobilized carrier that combines high loading capacity with long-term stability. The SiO₂@MnO₂@MAF-7 core–shell structured carrier was prepared via a solvothermal self-assembly method. Three immobilization strategies—adsorption, covalent cross-linking, and encapsulation—were systematically compared for their immobilization efficacy on HRP. The material structure was analyzed using techniques such as specific surface area analysis (BET), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) to characterize the material structure. Enzyme kinetic parameter determination experiments were conducted to systematically evaluate the performance advantages of the immobilized enzyme. BET analysis showed that SiO₂@MnO₂@MAF-7 had a specific surface area of 251.99 m²/g and a mesoporous area of 12.47 nm, and its HRP loading was 50.37 U/mg (immobilization efficiency 85.03%). Compared with free HRP, the Km value of the immobilized enzyme was decreased by 42%, the activity retention rate was increased by 35–50% at 80 °C and pH 4–9, and the activity was maintained by 65% after five repeated uses. In this study, MAF-7 was combined with MnO₂/SiO₂ for HRP immobilization for the first time, and the triple effect of rigid support-catalytic synergy-confined protection synergistically improved the stability of the enzyme, providing a new strategy for the industrial application of oxidoreductases.