An integrated microfluidic real-time PCR system for pathogen detection

The development of microfluidics-based PCR for rapid genetic analysis with direct application in pathogen detection is an emerging research field. System integration to effectively connect individual function modules is an important aspect for the practical use of microfluidics PCR for diagnosis applications. Here we present our preliminary development of a fully-integrated microfluidic Real-Time PCR system with thermal cycler and online fluorescence detection module. The PCR microfluidic chip was fabricated in PDMS for rapid prototyping. The thermal cycler allows rapid temperature cycling with high accuracy (i.e. the average heating and cooling rate are 4.46 degrees C/s and 4.36 degrees C/s respectively). Using this developed system, we successfully demonstrated real-time PCR detection of E. coli O157:H7 with 20 mu L of PCR reaction mixture. Further development will lead to a more compact and portable microfluidic Real Time PCR system that offers a useful tool for various biomedical research and clinical applications.