Electrochemical real-time DNA amplification and detection on a microchip

Abstract

In this work, we demonstrate the implementation of a recently developed electrochemical real-time polymerase chain reaction technique on a silicon microchip for simultaneous DNA amplification and detection. The real-time DNA amplification relies on an electroactive DNA intercalator, methylene blue (MB), for electrochemical signal measurements. Compared to other existing electrochemical methods for DNA amplicon analysis, our electrochemical detection method is more advantageous for no requiring probe-modified electrodes, or asymmetric PCR, or solide-phase PCR. Our reaction microchips provide micro PCR environment to promote the efficiency of DNA amplification since the reaction in the microchip wells cost low-volume solution (10μL) and rapid amplification time. In this study, we demonstrate the electrochemical relationship of MB and PCR amplicons with PCR cycles increase by amplifying a fragment of white spot syndrome virus (WSSV) genome DNA. The results of this study prove that the real-time DNA reaction microchip has a great convenience for the rapid, user-friendly, and low-volume real-time PCR assays in the direct detection of WSSV diagnosis.