Monolithic Centrifugal Microfluidic Platform for Bacteria Capture and Concentration, Lysis, Nucleic-Acid Amplification, and Real-Time Detection

We report the design, fabrication, and characterization of a polymer centrifugal microfluidic system for the specific detection of bacterial pathogens. This single-cartridge platform integrates bacteria capture and concentration, supernatant solution removal, lysis, and nucleic-acid sequence-based amplification (NASBA) in a single unit. The unit is fabricated using multilayer lamination and consists of five different polymer layers. Bacteria capture and concentration are accomplished by sedimentation in five minutes. Centrifugation forces also drive the subsequent steps. A wax valve is integrated in the cartridge to enable high-speed centrifugation. Oil is used to prevent evaporation during reactions requiring thermal cycling. Device functionality was demonstrated by real-time detection of E. coli from a 200-muL sample.