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Applied and Environmental Microbiology, March 2004, p. 1651-1657, Vol. 70, No. 3
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.3.1651-1657.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Rapid Detection and Enumeration of Legionella pneumophila in Hot Water Systems by Solid-Phase Cytometry

Helena Aurell,1,2 Philippe Catala,2 Pierre Farge,1 France Wallet,3 Matthieu Le Brun,3 Jürgen H. Helbig,4 Sophie Jarraud,1 and Philippe Lebaron2*

Centre National de Référence des Legionella, INSERM E-0230, Laboratoire de Bactériologie, Faculté de Médecine Laennec IFR 62, 69372 Lyon,1 Laboratoire Arago, Observatoire Océanologique, Centre National de la Recherche Scientifique (CNRS-UMR7621), Institut National des Sciences de l'Univers, Université Paris VI, 66651 Banyuls-sur-Mer,2 Electricité de France, Division Recherche et Développement, 78401 Chatou, France,3 Technical University Dresden, Institute of Medical Microbiology and Hygiene, D-01307 Dresden, Germany4

Received 11 September 2003/ Accepted 21 November 2003

A new method for the rapid and sensitive detection of Legionella pneumophila in hot water systems has been developed. The method is based on an IF assay combined with detection by solid-phase cytometry. This method allowed the enumeration of L. pneumophila serogroup 1 and L. pneumophila serogroups 2 to 6, 8 to 10, and 12 to 15 in tap water samples within 3 to 4 h. The sensitivity of the method was between 10 and 100 bacteria per liter and was principally limited by the filtration capacity of membranes. The specificity of the antibody was evaluated against 15 non-Legionella strains, and no cross-reactivity was observed. When the method was applied to natural waters, direct counts of L. pneumophila were compared with the number of CFU obtained by the standard culture method. Direct counts were always higher than culturable counts, and the ratio between the two methods ranged from 1.4 to 325. Solid-phase cytometry offers a fast and sensitive alternative to the culture method for L. pneumophila screening in hot water systems.

* Corresponding author. Mailing address: Laboratoire Arago, Observatoire Océanologique, Centre National de la Recherche Scientifique (CNRS-UMR7621), Institut National des Sciences de l'Univers, Université Paris VI, 66651 Banyuls-sur-Mer, France. Phone: 33 468 887 353. Fax: 33 468 887 398. E-mail: lebaron{at}obs-banyuls.fr.

Applied and Environmental Microbiology, March 2004, p. 1651-1657, Vol. 70, No. 3
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.3.1651-1657.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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