Semi-real time electrochemical monitoring for influenza virus RNA by reverse transcription loop-mediated isothermal amplification using a USB powered portable potentiostat

Abstract

In this paper, the semi-real time electrochemical monitoring method using a screen-printed electrode, which employs reverse transcription loop-mediated isothermal amplification (RT-LAMP) for influenza virus RNA, is presented. The amplified DNA combined with methylene blue (MB), which was used as an electroactive DNA intercalator, and the electrochemical signal was monitored using square wave voltammetry in the presence of RT-LAMP reagent components. MB molecules binding to amplified DNA caused the reduction of the peak current due to the slow diffusion of MB-amplified DNA complex to the electrode surface. We successfully monitored the amplification process of DNA on the basis of RT-LAMP by measuring and analyzing the electrochemical signal of MB with only one screen-printed electrode that connected with a USB powered portable potentiostat. The peak height of the current was related to the extent of amplification of DNA and the amount of input RNA. Since laborious probe immobilization is not required and both the amplification and the monitoring are possible in a single tube, our method does not suffer from potential cross-contamination. Furthermore, our method provides a new rote for the development of electrochemical hand held biosensors.

Future developments in biosensors for field-ready Zika virus diagnostics

“…4b,c). Many real-time nucleic acid quantification methods have also been used, though again not all for ZIKV, and include measurement of fluorescence [51], Mg + pyrophosphate [52], electrochemical [53], or colorimetric signal changes, detectable by the human eye [54, 55] or optical sensors [56–58]. …”

Field-deployable rapid multiple biosensing system for detection of chemical and biological warfare agents

“…Our research group has also reported a colorimetric sensor for the detection of DZO based on the inhibition of AChE 28 . Additionally, we have developed a portable electrochemical detection method using a USB-powered potentiostat and a disposable SPCE [19][20][21] . With these, we integrated our electrochemical biosensing device for chemical agent detection due to its portability and mobility.…”

“…Electrochemical measurement is desired due to its highly sensitive detection and, more importantly, the portability and miniaturization of capable devices. We have already developed a USB-powered portable potentiostat (BDTminiSTAT100) and a disposable screen-printed carbon electrode (SPCE) with exemplary performance [19][20][21] . The optimized electrochemical measurements from our devices were selected and then utilized in the developed autonomous system.…”

An Electrochemical DNA Biosensor Based on Au-reduced Graphene Oxide Nanocomposite for Transgenic Event Bt63 Detection

“…Electrochemical measurements were carried out using a USB powered portable potentiostat (BDTminiSTAT100), which was recommended as being detailed in the research before. 22 The planar screen-printed electrode (SPE) was purchased from BioDevice Technology Co., Ltd. (Japan), which consisted of two gold electrodes as the working and counter electrodes, respectively; an Ag/AgCl electrode was used as the reference electrode.…”