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Journal of Clinical Microbiology, January 2003, p. 368-372, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.368-372.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Attempted Isolation of Nanobacterium sp. Microorganisms from Upper Urinary Tract Stones

Michel Drancourt,1 Véronique Jacomo,1 Hubert Lépidi,2 Eric Lechevallier,3 Vincent Grisoni,4 Christian Coulange,3 Edith Ragni,5 Claude Alasia,6 Bertrand Dussol,7 Yvon Berland,7 and Didier Raoult1*

Unité des Rickettsies, CNRS: UPRESA 6020, IFR48,1 Service de Microscopie Électronique, Faculté de Médecine, Université de la Méditerranée,6 Service d'Urologie et Transplantation Rénale,2 Service d'Urologie, Hôpital Salvator,3 Service d'Urologie, Hôpital Nord,4 Laboratoire d'Anatomopathologie, Hôpital de la Timone,5 Service de Néphrologie et d'Hémodialyse, Hôpital Sainte-Marguerite, Marseille, France7

Received 14 January 2002/ Returned for modification 17 May 2002/ Accepted 19 June 2002

A single team has reported isolation of nanobacteria in human and bovine blood products, as well as, more recently, kidney stones. This has raised controversy. To confirm the data, we searched for nanobacteria from 10 aseptically removed upper urinary tract (UUT) stones. We used scanning electronic microscopy (SEM) with four stones and culture of stones on either 3T6 fibroblast monolayers or liquid RPMI medium. Detection of nanobacteria was made with a commercially available monoclonal antibody, 16S ribosomal DNA amplification with specific primers, and transmission electronic microscopy (TEM) of inoculated cells. SEM showed nanoparticles in four of four UUT stones similar to those recently described. TEM of inoculated 3T6 cell monolayers has shown transient intracytoplasmic vacuolar formations containing 200- to 500-nm particles in 3 of 10 cell cultures. Gimenez staining, Hoechst staining, and specific monoclonal immunofluorescence failed to reveal nanobacteria. Finally, we could not grow Nanobacterium sp. microorganisms by the techniques described. Although with SEM, we observed nanoparticles morphologically similar to nanobacteria, we failed to isolate Nanobacterium sp. microorganisms in culture and to prove the bacterial nature of these nanoparticles in stones.

* Corresponding author. Mailing address: Unité des Rickettsies, Faculté de Médecine, 27 Boulevard Jean Moulin, 13385 Marseille Cedex 5, France. Phone: 33 (0)4 91 83 55 17. Fax: 33 (0)4 91 83 03 90. E-mail: Didier.Raoult{at}medecine.univ-mrs.fr.

Journal of Clinical Microbiology, January 2003, p. 368-372, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.368-372.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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