A miniaturized quantitative polymerase chain reaction system for DNA amplification and detection

In this study, a new polymerase chain reaction (PCR) system incorporated with a fluorescence detection module and a micro PCR chip has been demonstrated to allow for on-line detection of infectious diseases. A new flow-through PCR chip was developed to reduce cooling and heating times for a PCR process. It is comprised of two micro modules for thermal and microfluidic control. The microfluidic control module in this chip adopts three serpentine-shape micropumps to rapidly transport DNA samples and PCR reagents through three reaction chambers. Using this approach, one can randomly adjust cycle numbers and reaction times of the DNA samples for each reaction, thus optimizing a PCR process. The micro thermal control module consists of three individual array-type heaters and temperature sensors to modulate three specific temperatures for three thermal steps of a PCR process. The optical detection system is then used for detection and quantification of the amplified products. The amplification and detection of detection genes associated with two viruses, specifically hepatitis B virus (HBV) and hepatitis C virus (HCV), has been performed to demonstrate the capabilities of the developed system. The miniaturized PCR system has been successfully used for DNA amplification and quantification and is a promising new tool for molecular diagnosis.