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Journal of Clinical Microbiology, October 2006, p. 3760-3767, Vol. 44, No. 10
0095-1137/06/$08.00+0     doi:10.1128/JCM.01435-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Evaluation of Peptide Nucleic Acid-Fluorescence In Situ Hybridization for Identification of Clinically Relevant Mycobacteria in Clinical Specimens and Tissue Sections

Michael Lefmann,^1, Birgitta Schweickert,¹ Petra Buchholz,¹ Ulf B. Göbel,¹ Timo Ulrichs,^2, Peter Seiler,^2, Dirk Theegarten,^3,¶ and Annette Moter^1*

Institut für Mikrobiologie und Hygiene, Charité—Universitätsmedizin Berlin, Dorotheenstrasse 96, 10117 Berlin, Germany,¹ Abteilung für Immunologie, Max-Planck-Institut für Infektionsbiologie, Schumannstrasse 21/22, 10117 Berlin, Germany,² Abteilung für Pathologie, Ruhr-Universität Bochum, Universitätsstrasse 150, 44780 Bochum, Germany³

Received 11 July 2006/ Accepted 5 August 2006

With fluorescently labeled PNA (peptide nucleic acid) probes targeting 16S rRNA, we established a 3-h fluorescence in situ hybridization (FISH) procedure for specific visualization of members of the Mycobacterium tuberculosis complex, M. leprae, M. avium, and M. kansasii. Probe specificity was tested against a panel of 25 Mycobacterium spp. and 10 gram-positive organisms. After validation, probes were used to identify 52 mycobacterial culture isolates. Results were compared to conventional genotypic identification with amplification-based methods. All isolates (M. tuberculosis complex, n = 24; M. avium, n = 7; M. kansasii, n = 1) were correctly identified by FISH. In addition, the technique was used successfully for visualization of mycobacteria in biopsies from infected humans or animals. In conclusion, PNA-FISH is a fast and accurate tool for species-specific identification of culture-grown mycobacteria and for direct visualization of these organisms in tissue sections. It may be used successfully for both research and clinical microbiology.

Present address: HAEMA AG, Institut für Medizinische Mikrobiologie am HELIOS-Klinikum Berlin-Buch, 13125 Berlin, Germany.

Present address: German Federal Ministry of Health, 10117 Berlin, Germany.

Present address: Actelion Pharmaceuticals Ltd., CH-4123 Allschwil, Switzerland.

^¶ Present address: Institut für Pathologie und Neuropathologie, Universitätsklinikum Essen, D-45122 Essen, Germany.