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Applied and Environmental Microbiology, April 2006, p. 2801-2808, Vol. 72, No. 4
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.4.2801-2808.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Quantitative Real-Time Legionella PCR for Environmental Water Samples: Data Interpretation

Philippe Joly,1 Pierre-Alain Falconnet,2 Janine André,3 Nicole Weill,3 Monique Reyrolle,1 François Vandenesch,1 Max Maurin,2 Jerome Etienne,1 and Sophie Jarraud1*

Centre National de Référence des Legionella, INSERM E-0230, Université de Lyon, Faculté de Médecine, IFR 62, 7 rue Guillaume-Paradin, 69372 Lyon Cedex 08, France,1 Laboratoire de Bactériologie du CHU de Grenoble, BP 217, 38043 Grenoble Cedex 09, France,2 CARSO-Laboratoire Santé Environnement Hygiène de Lyon (CARSO-LSEHL), 321 avenue Jean-Jaurès, 69362 Lyon Cedex 07, France3

Received 31 October 2005/ Accepted 6 February 2006

Quantitative Legionella PCRs targeting the 16S rRNA gene (specific for the genus Legionella) and the mip gene (specific for the species Legionella pneumophila) were applied to a total of 223 hot water system samples (131 in one laboratory and 92 in another laboratory) and 37 cooling tower samples (all in the same laboratory). The PCR results were compared with those of conventional culture. 16S rRNA gene PCR results were nonquantifiable for 2.8% of cooling tower samples and up to 39.1% of hot water system samples, and this was highly predictive of Legionella CFU counts below 250/liter. PCR cutoff values for identifying hot water system samples containing >103 CFU/liter legionellae were determined separately in each laboratory. The cutoffs differed widely between the laboratories and had sensitivities from 87.7 to 92.9% and specificities from 77.3 to 96.5%. The best specificity was obtained with mip PCR. PCR cutoffs could not be determined for cooling tower samples, as the results were highly variable and often high for culture-negative samples. Thus, quantitative Legionella PCR appears to be applicable to samples from hot water systems, but the positivity cutoff has to be determined in each laboratory.

* Corresponding author. Mailing address: Centre National de Référence des Legionella, INSERM E-0230, Faculté de Médecine, IFR 62, 7 rue Guillaume-Paradin, 69372 Lyon Cedex 08, France. Phone: (33) 47 877 87 26. Fax: (33) 47 877 86 58. E-mail: sophie.jarraud{at}univ-lyon1.fr.

Applied and Environmental Microbiology, April 2006, p. 2801-2808, Vol. 72, No. 4
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.4.2801-2808.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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