An unconventional approach to impedance microbiology: detection of culture media conductivity variations due to bacteriophage generated lyses of host bacteria

Biosens Bioelectron. 2015 May 15:67:615-20. doi: 10.1016/j.bios.2014.09.075. Epub 2014 Sep 30.

Abstract

A novel and unconventional approach to impedance microbiology has been under investigation. In our approach, solution conductivity variations are generated from bacteriophage lyses of infected host cells and the consequent release of conductive endoplasmic material. To sensitively detect the lysis, low conductive growth media have been developed. A microchip has been fabricated to perform the analysis. The microchip is made of two bare gold electrodes and PDMS microchamber of 36 nL volume. Escherichia coli and selective phages T4 have been used as case study. Proof-of-principle experiments are here presented and discussed. The method was characterised in a wide range between 10(4) and 10(8) CFU/mL, where linear relation was found between conductivity variation and cell concentration in a log10 vs. log10 plot. The method is suited to integration with sample preparation based on phage-functionalised magnetic beads. It has a potential detection limit below 1 CFU/chamber and a total assay time of less than 1 h.

Keywords: Bacteriophage; Impedance microbiology; Low conductivity broth; Microsystem; Pathogen detection; System integration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophage T4 / growth & development
  • Bacteriophage T4 / isolation & purification*
  • Biosensing Techniques*
  • Culture Media / analysis*
  • Culture Media / chemistry
  • Electric Conductivity
  • Electric Impedance
  • Escherichia coli / isolation & purification*
  • Escherichia coli / virology
  • Gold / chemistry
  • Microchip Analytical Procedures

Substances

  • Culture Media
  • Gold