Evaluation of peptide nucleic acid-fluorescence in situ hybridization for identification of clinically relevant mycobacteria in clinical specimens and tissue sections.
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.
Lefmann
Michael
M
Institut für Mikrobiologie und Hygiene, Charité-Universitätsmedizin Berlin, Dorotheenstr. 96, 10117 Berlin, Germany. annette.moter@charite.de.
Schweickert
Birgitta
B
Buchholz
Petra
P
Göbel
Ulf B
UB
Ulrichs
Timo
T
Seiler
Peter
P
Theegarten
Dirk
D
Moter
Annette
A
D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D009161:Mycobacterium / Q000145:classification* / Q000302:isolation & purification; D009164:Mycobacterium Infections / Q000175:diagnosis* / Q000382:microbiology; D015341:Nucleic Acid Probes; D020135:Peptide Nucleic Acids; D012680:Sensitivity and Specificity
D015341:Nucleic Acid Probes; D020135:Peptide Nucleic Acids