High-throughput centrifugal microfluidic platform for multiplex respiratory virus diagnostics

We developed an advanced point-of-care (POC) molecular diagnostic platform that utilizes a centrifugal microfluidic chip for automating the diagnostics process of prevalent respiratory viruses, including Influenza A (H1N1, H3N2), Influenza B, and SARS-CoV-2, in a high-throughput (HTP) manner. The proposed POC microsystem can perform not only the whole processes including RNA purification, the reverse transcription loop-mediated isothermal amplification (RT-LAMP), and the real-time fluorescence detection, but also 30 sample analysis simultaneously in one run. Owing to the innovative zigzag aliquoting structure on the centrifugal microfluidic chip, the ultra-rapid and equivalent reagent division into 30 chambers with a single shot injection were possible, which expedited the high-throughput RNA extraction. The purified RNA templates were mixed with lyophilized RT-LAMP reagents and directed to the reaction chambers, where the primer sets for each target respiratory virus were freeze-dried for isothermal amplification. A total of 150 reactions (5 reaction chambers/unit × 30 units) were carried out on a single chip, so multiple and high-throughput diagnostics for 30 clinical samples could be completed in 1.5 hrs. Our platform with the high-throughput screening capability could be operated automatically, making it an invaluable tool for swift responses to viral pandemics such as COVID-19.