Integrated biochip for label-free and real-time detection of DNA amplification by contactless impedance measurements based on interdigitated electrodes

Here, we introduce an integrated biochip which offers accurate thermal control and sensitive electrochemical detection of DNA amplification in real-time. The biochip includes a 10 mu l microchamber, a temperature sensor, a heater, and a contactless impedance biosensor. A pair of interdigitated electrodes is employed as the impedance biosensor and the products of the amplification are determined directly through tracing the impedance change, without using any labels, redox indicators, or probes. Real-time monitoring of strand-displacement amplification and rolling circle amplification was successfully performed on the biochip and a detection limit of 1 nM was achieved. Amplification starting at an initial concentration of 10 nM could be discriminated from that starting at 1 nM started concentration as well as from the negative control. Since an insulation layer covers the electrodes, the electrodes are spared from erosion, hydrolysis and bubble formation on the surface, thus, ensuring a long lifetime and a high reusability of the sensor. In comparison to bench-top apparatus, our chip shows good efficiency, sensitivity, accuracy, and versatility. Our system requires only simple equipments and simple skills, and can easily be miniaturized into a micro-scale system. The system will then be suitable for a handheld portable device, which can be applied in remote areas. It covers merits such as low cost, low-power consumption, rapid response, real-time monitoring, label-free detection, and high-throughput detection. (c) 2013 Elsevier B.V. All rights reserved.