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 influenzavirus RNA, is presented. The amplified DNA combined with methylene blue (MB), which was used asan electroactive DNA intercalator, and the electrochemical signal was monitored using square wavevoltammetry in the presence of RT-LAMP reagent components. MB molecules binding to amplifiedDNA caused the reduction of the peak current due to the slow diffusion of MB-amplified DNAcomplex to the electrode surface. We successfully monitored the amplification process of DNA on thebasis of RT-LAMP by measuring and analyzing the electrochemical signal of MB with only one screenprintedelectrode that connected with a USB powered portable potentiostat. The peak height of thecurrent was related to the extent of amplification of DNA and the amount of input RNA. Sincelaborious probe immobilization is not required and both the amplification and the monitoring arepossible 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.
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