Evaluation of propidium monoazide (PMA) treatment directly on membrane filter for the enumeration of viable but non cultivable Legionella by qPCR

J Microbiol Methods. 2012 Feb;88(2):319-21. doi: 10.1016/j.mimet.2011.12.010. Epub 2011 Dec 22.

Abstract

A PMA (propidium monoazide) pretreatment protocol, in which PMA is applied directly to membrane filters, was developed for the PCR-based quantification (PMA-qPCR) of viable Legionella pneumophila. Using this method, the amplification of DNA from membrane-damaged L. pneumophila was strongly inhibited for samples containing a small number of dead bacteria.

MeSH terms

  • Azides / chemistry*
  • Bacteriological Techniques / methods*
  • DNA, Bacterial / analysis
  • Legionella / cytology
  • Legionella / genetics
  • Legionella / isolation & purification*
  • Membranes, Artificial
  • Microbial Viability
  • Polymerase Chain Reaction / methods*
  • Propidium / analogs & derivatives*
  • Propidium / chemistry

Substances

  • Azides
  • DNA, Bacterial
  • Membranes, Artificial
  • propidium monoazide
  • Propidium