Simultaneous electrochemical magneto genosensing of foodborne bacteria based on triple-tagging multiplex amplification

Biosens Bioelectron. 2015 Dec 15:74:652-9. doi: 10.1016/j.bios.2015.07.008. Epub 2015 Jul 9.

Abstract

Simultaneous detection of Salmonella enterica, Listeria monocytogenes and Escherichia coli based on triple-tagging multiplex PCR and electrochemical magneto genosensing on silica magnetic particles is reported. A set of tagging primers were selected for the specific amplification of yfiR (375 bp), hlyA (234 bp) and eaeA (151bp), being one of the primers for each set labelled with fluorescein, biotin and digoxigenin coding for S. enterica, L. monocytogenes and E. coli, respectively. Afterwards, electrochemical magneto genosensing of the bacteria was achieved by using silica magnetic particles as a carrier and three different electrochemical reporters, specific for each pathogen. This method was able to clearly distinguish among the pathogenic bacteria tested within 50 min, with detection limits ranging from 12 to 46 pg μL(-1).

Keywords: Electrochemical genosensing; Foodborne bacteria; Magneto-actuated electrode; Simultaneous detection; Triple-tagging multiplex PCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Load / instrumentation*
  • Biosensing Techniques / instrumentation
  • Complex Mixtures / analysis
  • Conductometry / instrumentation*
  • Equipment Design
  • Equipment Failure Analysis
  • Food Analysis / instrumentation*
  • Food Contamination / analysis*
  • Food Microbiology / instrumentation*
  • Magnetite Nanoparticles / chemistry
  • Magnetite Nanoparticles / ultrastructure
  • Magnetometry / instrumentation*
  • Micro-Electrical-Mechanical Systems / instrumentation
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Silicon Dioxide / chemistry
  • Systems Integration

Substances

  • Complex Mixtures
  • Magnetite Nanoparticles
  • Silicon Dioxide