Development of a microfluidic isothermal amplification nucleic acid detection (mFIND) platform for simultaneous
Peiyan He1, Yating Feng2, Xiaojie Zhou2
1Jiaxing Center for Disease Control and Prevention, No. 486, Wen Qiao Road, Jiaxing, Zhejiang, China.
Abstract:
We developed the mFIND platform, a centrifugal microfluidic isothermal amplification system for simultaneous detection of influenza A/B (FLUAV/FLUBV), SARS-CoV-2, and RSV - key agents of acute respiratory infections. The chip integrates four independent reaction chambers preloaded with lyophilized reagents, enabling room-temperature storage and a simplified two-step workflow. A portable analyzer automates sample metering, isothermal amplification (41 ± 0.4 °C), and real-time fluorescence monitoring, completing detection within 15 min with an ultra-low reaction volume of 2.5 μL per chamber. The platform achieved limits of detection of 2 copies/test for FLUAV/FLUBV and 5 copies/test for SARS-CoV-2/RSV, with no cross-reactivity against 23 non-target pathogens. Clinical validation using 236 positive samples showed 100% positive and negative agreement with qPCR. The mFIND platform combines rapid turnaround, simplified workflow, low reagent consumption, and parallel processing of four samples (16 targets), positioning it as a practical point-of-care nucleic acid testing solution for decentralized settings.
