Efficacy of the detection of Legionella in hot and cold water samples by culture and PCR. I. Standardization of methods
Journal Title: Annals of Agricultural and Environmental Medicine - Year 2012, Vol 19, Issue 2
Abstract
The aim of the present study was: – to compare methods for concentration and isolation of [i]Legionella[/i] DNA from water; – to examine the efficacy of various modifications of PCR test (PCR, semi-nested PCR, and real-time PCR) for the detection of known numbers of [i]Legionella pneumophila[/i] in water samples artificially contaminated with the strain of this bacterium and in randomly selected samples of environmental water, in parallel with examination by culture. It was found that filtration is much more effective than centrifugation for the concentration of DNA in water samples, and that the Qiamp DNA Mini-Kit is the most efficient for isolation of Legionella DNA from water. The semi-nested PCR and real-time PCR proved to be the most sensitive methods for detection of [i]Legionella[/i] DNA in water samples. Both PCR modifications showed a high correlation with recovery of [i]Legionella[/i] by culture (p<0.01), while no correlation occurred between the results of one-stage PCR and culture (p>0.1).
Authors and Affiliations
Angelina Wójcik-Fatla, Nimfa Maria Stojek, Jacek Dutkiewicz
Keywords
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