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Journal of Clinical Microbiology, December 2004, p. 5493-5501, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5493-5501.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Amoebal Coculture of "Mycobacterium massiliense" sp. nov. from the Sputum of a Patient with Hemoptoic Pneumonia

Toïdi Adékambi,1 Martine Reynaud-Gaubert,2 Gilbert Greub,1,3 Marie-José Gevaudan,1 Bernard La Scola,1 Didier Raoult,1 and Michel Drancourt1*

Unité des Rickettsies, CNRS UMR 6020 IFR 48, Faculté de Médecine, Université de la Méditerranée,1 Service des Maladies Respiratoires, Hôpital Sainte Marguerite, Marseille, France,2 Microbiology Institute, Faculty of Biology and Medicine, University of Lausanne, Lausanne, Switzerland3

Received 15 April 2004/ Returned for modification 21 July 2004/ Accepted 10 August 2004

A nonphotochromogenic, rapidly growing Mycobacterium strain was isolated in pure culture from the sputum and the bronchoalveolar fluid of a patient with hemoptoic pneumonia by using axenic media and an amoebal coculture system. Both isolates grew in less than 7 days at 24 to 37°C with an optimal growth temperature of 30°C. The isolates exhibited biochemical and antimicrobial susceptibility profiles overlapping those of Mycobacterium abscessus, Mycobacterium chelonae, and Mycobacterium immunogenum, indicating that they belonged to M. chelonae-M. abscessus group. They differed from M. abscessus in ß-galactosidase, ß-N-acetyl-ß-glucosaminidase, and ß-glucuronidase activities and by the lack of nitrate reductase and indole production activities, as well as in their in vitro susceptibilities to minocycline and doxycycline. These isolates and M. abscessus differed from M. chelonae and M. immunogenum by exhibiting gelatinase and tryptophane desaminase activities. Their 16S rRNA genes had complete sequence identity with that of M. abscessus and >99.6% similarity with those of M. chelonae and M. immunogenum. Further molecular investigations showed that partial hsp65 and sodA gene sequences differed from that of M. abscessus by five and three positions over 441 bp, respectively. Partial rpoB and recA gene sequence analyses showed 96 and 98% similarities with M. abscessus, respectively. Similarly, 16S-23S rRNA internal transcribed spacer sequence of the isolates differed from that of M. abscessus by a A->G substitution at position 60 and a C insertion at position 102. Phenotypic and genotypic features of these two isolates indicated that they were representative of a new mycobacterial species within the M. chelonae-M. abscessus group. Phylogenetic analysis suggested that these isolates were perhaps recently derived from M. abscessus. We propose the name of "Mycobacterium massiliense" for this new species. The type strain has been deposited in the Collection Institut Pasteur as CIP 108297T and in Culture Collection of the University of Göteborg, Göteborg, Sweden, as CCUG 48898T.

* Corresponding author. Mailing address: Unité des Rickettsies, Faculté de Médecine, 27 Boulevard Jean Moulin, 13385 Marseille Cedex 05, France. Phone: (33) 04-91-32-43-75. Fax: (33) 04-91-38-77-72. E-mail: Michel.Drancourt{at}medecine.univ-mrs.fr.

Journal of Clinical Microbiology, December 2004, p. 5493-5501, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5493-5501.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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