Fluidic handling system for PCR-based sample-to-answer detection of viral nucleic acids

Abstract

Sample-to-answer detection of a viral infectious disease is becoming increasingly important to prevent pandemic outbreaks. However, only a few microfluidic chips that use nucleic acid isolation and detection have implemented all-in-one assays in a simple and effective platform. Here, we present a microfluidic platform that isolates and detects viral RNA of influenza A (H1N1) virus from clinical swab samples in a fully automated, effective, and highly sensitive manner. The device with a vertically movable top layer and a rotationally mobile bottom layer exploits surface tension in a solution. In this way, the device effectively orchestrates multiple functions, including transporting, mixing, and washing of reagents and magnetic beads without the need for on-chip valves, pumps, and microfluidic networks that increase operational and structural complexity. With integrated heating and detection functions, the device provides fully automated sample-to-answer detection with real-time polymerase chain reaction in 60 min for positive results with high sensitivity with the limit of detection of 26 copies/reaction. Our platform is useful for accurate diagnosis of infectious diseases in point-of-care settings.