Ultra-fast electronic detection of antimicrobial resistance genes using isothermal amplification and Thin Film Transistor sensors

Biosens Bioelectron. 2017 Oct 15:96:281-287. doi: 10.1016/j.bios.2017.05.016. Epub 2017 May 9.

Abstract

A low cost thin-film transistor (TFT) nanoribbon (NR) sensor has been developed for rapid real-time detection of DNA amplification using an isothermal Recombinase Polymerase Amplification (RPA) method. The semiconductor chip measures DNA amplification through a pH change, rather than via fluorescence. The utility of the method was demonstrated by amplifying CTX-M and NDM, two genes that confer bacterial resistance to cephalosporins and carbapenems, respectively. It is shown that this approach provides extremely fast and sensitive detection. It can detect <10 copies of the gene in genomic DNA extracted from E. coli or K. pneumoniae clinical isolates within a few minutes. A differential readout system was developed to minimize the effect of primer-dimer amplification on the assay. The simple device has the potential for low cost, portable and real-time nucleic acid analysis as a Point of Care device.

Keywords: Antimicrobial resistance (AMR); Isothermal nucleic acid amplification; Recombinase Polymerase Amplification (RPA); Thin Film Transistor (TFT).

MeSH terms

  • Biosensing Techniques / instrumentation*
  • DNA, Bacterial / analysis
  • DNA, Bacterial / genetics*
  • Drug Resistance, Bacterial
  • Equipment Design
  • Escherichia coli / genetics*
  • Escherichia coli Infections / microbiology
  • Genes, Bacterial*
  • Humans
  • Klebsiella Infections / microbiology
  • Klebsiella pneumoniae / genetics*
  • Nucleic Acid Amplification Techniques / instrumentation*
  • Semiconductors
  • Transistors, Electronic*

Substances

  • DNA, Bacterial