11:30 AM - 11:45 AM
▲ [13a-N322-10] Real-time PCR detection with electrochemical impedance spectroscopy
Keywords:EIS, real-time PCR
The Polymerase chain reaction is a primary DNA amplification technique in molecular technology and provided the basis for rapid and reliable detection of pathogen, especially untiled real-time PCR for virus detection. Real-time PCR relied on fluorescent reporters. However, the fluorescent dyes are easily degraded in light, and problems with storage and reader lead to expensive systems and bulky instruments. In order to develop portable, cost effective and robust devices, electrochemical technology is attractive as PCR detection methods. Electrochemistry can detect target DNA by a change in the state of charge transfer at the electrode surface in a wide range of ionic solutions. The electrodes also could withstand the effect of thermal cycles. We developed a new electrochemical impedance method for real-time detection of PCR.
The [Ru(bpy)2DPPZ]2+ was a DNA intercalator that could intercalate into the double strained DNA (dsDNA). We found that [Ru(bpy)2DPPZ]2+ is not only a DNA intercalator, but it also had a special effect. The synthesized DNA intercalator [Ru(bpy)2DPPZ]2+ promoted charge transfer between hexacyanoferrate ions and a carbon electrode at only 1 µM, resulting in a significant decrease in impedance. This effect also could be observed in cyclic voltammetry (CV). We fabricated a protective layer of polyvinyl alcohol (PVA) on the GC electrode against non-specific adsorption then measured the EIS. The Rct with heat cycles showed an S curve, and the increased point clearly different from the background. This increase curve could be clearer with DRct (Figure 2). The results demonstrated that the combination of the Ru effect and the PVA coat electrode could monitor PCR progress in real-time.This research is partially supported by the COI (Grant No. JPMJCE1310), CREST (Grant No. JPMJCR18I3) from JST.
The [Ru(bpy)2DPPZ]2+ was a DNA intercalator that could intercalate into the double strained DNA (dsDNA). We found that [Ru(bpy)2DPPZ]2+ is not only a DNA intercalator, but it also had a special effect. The synthesized DNA intercalator [Ru(bpy)2DPPZ]2+ promoted charge transfer between hexacyanoferrate ions and a carbon electrode at only 1 µM, resulting in a significant decrease in impedance. This effect also could be observed in cyclic voltammetry (CV). We fabricated a protective layer of polyvinyl alcohol (PVA) on the GC electrode against non-specific adsorption then measured the EIS. The Rct with heat cycles showed an S curve, and the increased point clearly different from the background. This increase curve could be clearer with DRct (Figure 2). The results demonstrated that the combination of the Ru effect and the PVA coat electrode could monitor PCR progress in real-time.This research is partially supported by the COI (Grant No. JPMJCE1310), CREST (Grant No. JPMJCR18I3) from JST.