AEM
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Pougnard, C.
Right arrow Articles by Lebaron, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Pougnard, C.
Right arrow Articles by Lebaron, P.
Agricola
Right arrow Articles by Pougnard, C.
Right arrow Articles by Lebaron, P.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, June 2002, p. 3102-3107, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.3102-3107.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Rapid Detection and Enumeration of Naegleria fowleri in Surface Waters by Solid-Phase Cytometry

Claire Pougnard,1 Philippe Catala,2 Jean-Louis Drocourt,3 Stephane Legastelois,4 Pierre Pernin,5 Emmanuelle Pringuez,1 and Philippe Lebaron2*

Electricité de France, Division Recherche et Développement, F-78401 Chatou Cedex,1 Observatoire Océanologique, Centre National de la Recherche Scientifique (CNRS-UMR7621), Institut National des Sciences de l'Univers, Université Paris VI, F-66651 Banyuls-sur-Mer Cedex,2 Chemunex, F-94854 Ivry-sur-Seine Cedex,3 Indicia Biotechnology, F-69600 Oullins,4 Laboratoire de Biologie Cellulaire EA-1655, Faculté de Médecine et de Pharmacie, F-69373 Lyon, France5

Received 28 December 2001/ Accepted 3 April 2002

A new method for the rapid and accurate detection of pathogenic Naegleria fowleri amoebae in surface environmental water was developed. The method is based on an immunofluorescent assay combined with detection by solid-phase cytometry. In this study we developed and compared two protocols using different reporter systems conjugated to antibodies. The monoclonal antibody Ac5D12 was conjugated with biotin and horseradish peroxidase, and the presence of cells was revealed with streptavidin conjugated to both R-phycoerythrin and cyanine Cy5 (RPE-Cy5) and tyramide-fluorescein isothiocyanate, respectively. The RPE-Cy5 protocol was the most efficient protocol and allowed the detection of both trophozoite and cyst forms in water. The direct counts obtained by this new method were not significantly different from those obtained by the traditional culture approach, and results were provided within 3 h. The sensitivity of the quantitative method is 200 cells per liter. The limit is due only to the filtration capacity of the membrane used.

* Corresponding author. Mailing address: Observatoire Océanologique, Microbiologie de l'Environnement, BP44, F-66651 Banyuls-sur-Mer Cedex, France. Phone: (33)4-68-88-73-53. Fax: (33)4-68-88-73-98. E-mail: lebaron{at}obs-banyuls.fr.

Applied and Environmental Microbiology, June 2002, p. 3102-3107, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.3102-3107.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:



Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
J. Bacteriol. Microbiol. Mol. Biol. Rev. Eukaryot. Cell All ASM Journals

Copyright © 2002 by the American Society for Microbiology. All rights reserved.