Ion sensing (EIS) real-time quantitative monitorization of isothermal DNA amplification

Biosens Bioelectron. 2014 Feb 15:52:50-5. doi: 10.1016/j.bios.2013.08.029. Epub 2013 Aug 24.

Abstract

Field-effect-based devices are becoming a basic structural element in a new generation of microbiosensors. Reliable molecular characterization of DNA and/or RNA is of paramount importance for disease diagnostics and to follow up alterations in gene expression profiles. The use of such devices for point-of-need diagnostics has been hindered by the need of standard or real-time PCR amplification procedures. The present work focuses on the development of a tantalum pentoxide (Ta2O5) based sensor for the real-time label free detection of DNA amplification via loop mediated isothermal amplification (LAMP) allowing for quantitative analysis of the cMYC proto-oncogene. The strategy based on the field effect sensor was tested within a range of 1 × 10(8)-10(11) copies of target DNA, and a linear relationship between the log copy number of the initial template DNA and threshold time was observed allowing for a semi-quantitative analysis of DNA template. The concept offers many of the advantages of isothermal quantitative real-time DNA amplification in a label free approach and may pave the way to point-of-care quantitative molecular analysis focused on ease of use and low cost.

Keywords: Cancer; DNA; EIS; Ion sensitive field effect sensors; Isothermal amplification; LAMP; Label free; Real time; Tantalum pentoxide; c-Myc; qRT-PCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques / methods*
  • DNA / isolation & purification*
  • Nucleic Acid Amplification Techniques
  • Oxides / chemistry
  • RNA / isolation & purification*
  • Tantalum / chemistry

Substances

  • Oxides
  • RNA
  • Tantalum
  • DNA
  • tantalum oxide