Integrated Real-Time PCR for Detection and Monitoring of Legionella pneumophila in Water Systems

Author:

Yaradou Diaraf Farba1,Hallier-Soulier Sylvie2,Moreau Sophie3,Poty Florence3,Hillion Yves2,Reyrolle Monique1,André Janine4,Festoc Gabriel2,Delabre Karine3,Vandenesch François1,Etienne Jerome1,Jarraud Sophie1

Affiliation:

1. INSERM E0230, Lyon F-69008, France, and Université Lyon 1, Centre National de Référence des Legionella, Faculté Laennec, Lyon F-69008, France

2. GeneSystems, 1 rue du Courtil, Centre d'Affaires CICEA, Bâtiment 1, 35170 Bruz, France

3. CAE, Veolia Environnement, 1 place de Turenne, 94417 Saint Maurice, France

4. CARSO-Laboratoire Santé Environnement Hygiène de Lyon (CARSO-LSEHL), 321 avenue Jean-Jaurès, 69362 Lyon cedex 07, France

Abstract

ABSTRACT We evaluated a ready-to-use real-time quantitative Legionella pneumophila PCR assay system by testing 136 hot-water-system samples collected from 55 sites as well as 49 cooling tower samples collected from 20 different sites, in parallel with the standard culture method. The PCR assay was reproducible and suitable for routine quantification of L. pneumophila . An acceptable correlation between PCR and culture results was obtained for sanitary hot-water samples but not for cooling tower samples. We also monitored the same L. pneumophila -contaminated cooling tower for 13 months by analyzing 104 serial samples. The culture and PCR results were extremely variable over time, but the curves were similar. The differences between the PCR and culture results did not change over time and were not affected by regular biocide treatment. This ready-to-use PCR assay for L. pneumophila quantification could permit more timely disinfection of cooling towers.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Reference30 articles.

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2. Association Française de Normalisation. 2003. Water quality. Detection and enumeration of Legionella spp and Legionella pneumophila . Method by direct inoculation and after concentration by membrane filtration or centrifugation. AFNOR NF T90-431. http://www.boutique.afnor.fr/boutique.asp .

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5. Best, M., V. L. Yu, J. Stout, A. Goetz, R. R. Muder, and F. Taylor. 1983. Legionellaceae in the hospital water-supply. Epidemiological link with disease and evaluation of a method for control of nosocomial legionnaires' disease and Pittsburgh pneumonia. Lancetii:307-310.

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