A PCR microreactor machinery with passive micropump and battery-powered heater for thermo-cycled amplifications of clinical-level and multiplexed DNA targets

摘要

A self-contained polymerase chain reaction (PCR) platform with miniaturized power-system is introduced. Itis powered by portable lithium batteries and integrated continuous-flow PCR amplification platform. Generally speaking, traditional commercial thermal cyclers rely on external electric supply and thus they are too big in instrument size. This prevents real-timely and field testing during PCR diagnosis. The authors are introducing a continuous-flow 3D spiral microreactor for DNA amplifications and high-resolution multiplexed targets' detection by utilizing the polyvinyl chloride (PVC) tubing-polymer to fabricate the microreactor for the first time. The whole setup (that can all be placed in one hand) includes (a) the thermo-cycled control (5.5cm width, 10cm length and 11cm height), (b) the passive continuous-flow control, and (c) the trapezoidal PCR microreactor. The PCR platform can work for 4.5h continuously. With minimal accessories and operations, the total cost of the self-contained PCR machinery is 20 , much lower than the mainstream of commercial PCR machinery. By waiving external electric supply, this miniaturized PCR platform is applied to amplify the typical DNA fragments of plasma isolated hepatitis B virus (HBV), influenza virus (H7N9avian influenza) bacterium (Escherichia coli) plasmid and multiplexed targets. The efficiency of the method is 70% of that of commercial thermal cycler (CFX Connect, Bio Rad). The DNA of H7N9avian influenza can be detected in concentrations as low as 10(3) copies per L.By utilizing the polyvinyl chloride (PVC) tubing-polymer to fabricate the microchip for the first time, this paper introduces a 3D spiral microreactor with a miniaturized power-system supplied by portable AA-batteries, applied in DNA amplifications and high-resolution multiplexed targets' detection. In this microdevice, we made the machinery portable by waiving the external plugs, which solved the problem of traditional commercial thermal cyclers about large volume and expensive price mu L.