Discrimination of viable and dead fecal Bacteroidales bacteria by quantitative PCR with propidium monoazide

Appl Environ Microbiol. 2009 May;75(9):2940-4. doi: 10.1128/AEM.01333-08. Epub 2009 Mar 6.

Abstract

Propidium monoazide (PMA) was optimized to discriminate between viable and dead Bacteroides fragilis cells and extracellular DNA at different concentrations of solids using quantitative PCR. Conditions of 100 microM PMA and a 10-min light exposure also excluded DNA from heat-treated cells of nonculturable Bacteroidales in human feces and wastewater influent and effluent.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Azides / metabolism
  • Azides / pharmacology*
  • Bacteroidaceae / genetics
  • Bacteroidaceae / metabolism
  • Bacteroidaceae / physiology*
  • DNA / metabolism
  • Feces / microbiology
  • Fluorescent Dyes / metabolism
  • Fluorescent Dyes / pharmacology*
  • Microbial Viability*
  • Polymerase Chain Reaction / methods*
  • Propidium / analogs & derivatives*
  • Propidium / metabolism
  • Propidium / pharmacology
  • Spores / genetics
  • Spores / metabolism
  • Spores / physiology*
  • Water Microbiology

Substances

  • Azides
  • Fluorescent Dyes
  • propidium monoazide
  • Propidium
  • DNA