Quantitative real-time monitoring of RCA amplification of cancer biomarkers mediated by a flexible ion sensitive platform

Biosens Bioelectron. 2017 May 15:91:788-795. doi: 10.1016/j.bios.2017.01.052. Epub 2017 Jan 24.

Abstract

Ion sensitive field-effect transistors (ISFET) are the basis of radical new sensing approaches. Reliable molecular characterization of specific detection of DNA and/or RNA is vital for disease diagnostics and to follow up alterations in gene expression profiles. Devices and strategies for biomolecular recognition and detection should be developed into reliable and inexpensive platforms. Here, we describe the development of a flexible thin-film sensor for label free gene expression analysis. A charge modulated ISFET based sensor was integrated with real-time DNA/RNA isothermal nucleic acid amplification: Loop-mediated isothermal amplification (LAMP) and Rolling Circle Amplification (RCA) techniques for c-MYC and BCR-ABL1 genes, allowing for the real-time quantification of template. Also, RCA allowed the direct quantification of RNA targets at room temperature, eliminating the requirement for external temperature controllers and overall complexity of the molecular diagnostic approach. This integration between the biological and the sensor/electronic approaches enabled the development of an inexpensive and direct gene expression-profiling platform.

Keywords: Cancer; Cmisfet; Field-effect sensors; Gene expression analysis; Rca.

MeSH terms

  • Biomarkers, Tumor / genetics
  • Biosensing Techniques / instrumentation
  • Cell Line, Tumor
  • Electrodes
  • Equipment Design
  • Fusion Proteins, bcr-abl / genetics
  • Gene Expression Profiling / instrumentation*
  • Genes, myc
  • Humans
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / diagnosis
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics
  • Neoplasms / diagnosis
  • Neoplasms / genetics*
  • Nucleic Acid Amplification Techniques / instrumentation*

Substances

  • Biomarkers, Tumor
  • Fusion Proteins, bcr-abl