Centrifugal microfluidic platform for rapid PCR amplification using integrated thermoelectric heating and ice-valving

Abstract

We present a novel centrifugal microfluidic system for the rapid amplification of a Bacillus anthracis gene, with integrated pumping and thermoelectric-based heating and valving. Polymerase chain reaction is a powerful method for rapid and accurate diagnosis of infectious diseases and bio-warfare agents. The fully integrated system and disposable disc developed here automates the fluidic pumping, valving, and rapid thermocycling required for polymerase chain reaction. We demonstrate the first utilization of ice-valving in an integrated centrifugal microfluidic system as a method for sealing the thermocycling chamber to reduce fluid loss due to evaporation. The automated centrifugal system is validated via amplification of a B. anthracis gene. Fluidic thermal profiles are monitored and optimized to minimize non-specific amplification, reducing the total amplification time from 110 min down to 53 min.