A filter-based propidium monoazide technique to distinguish live from membrane-compromised microorganisms using quantitative PCR

Kristen N Hellein; Elizabeth M Kennedy; Valerie J Harwood; Katrina V Gordon; Shiao Y Wang; Joe Eugene Lepo

doi:10.1016/j.mimet.2012.01.015

A filter-based propidium monoazide technique to distinguish live from membrane-compromised microorganisms using quantitative PCR

J Microbiol Methods. 2012 Apr;89(1):76-8. doi: 10.1016/j.mimet.2012.01.015. Epub 2012 Jan 31.

Authors

Kristen N Hellein¹, Elizabeth M Kennedy, Valerie J Harwood, Katrina V Gordon, Shiao Y Wang, Joe Eugene Lepo

Affiliation

¹ Center for Environmental Diagnostics and Bioremediation, University of West Florida, 11000 University Parkway, Pensacola, FL 32514, USA.

PMID: 22314021
DOI: 10.1016/j.mimet.2012.01.015

Abstract

Propidium monoazide (PMA) was used to differentiate live from membrane-compromised bacteria in PCR methods. We have adapted this technique for use on membrane-filtered water samples and determined its efficacy using qPCR. Independent labs at three institutions replicated these findings.

Publication types

Evaluation Study
Multicenter Study
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Azides / metabolism*
Bacteriological Techniques / methods*
Cell Membrane / physiology*
Enzyme Inhibitors / metabolism*
Filtration / methods
Microbial Viability*
Propidium / analogs & derivatives*
Propidium / metabolism
Real-Time Polymerase Chain Reaction / methods*
Reproducibility of Results
Water Microbiology

Substances

Azides
Enzyme Inhibitors
propidium monoazide
Propidium