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Journal of Clinical Microbiology, September 1999, p. 2760-2765, Vol. 37, No. 9
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Fluorescence In Situ Hybridization Assay Using Peptide Nucleic Acid Probes for Differentiation between Tuberculous and Nontuberculous Mycobacterium Species in Smears of Mycobacterium Cultures

Henrik Stender,1 Kaare Lund,1 Kenneth H. Petersen,1 Ole F. Rasmussen,1,* Poonpilas Hongmanee,2 Håkan Miörner,3 and Sven E. Godtfredsen1

DAKO A/S, 2600 Glostrup,1 and Department of Mycobacteriology, Statens Serum Institut, 2300 Copenhagen,3 Denmark, and Department of Pathology, Ramathibodi Hospital, 10400 Bangkok, Thailand2

Received 8 March 1999/Returned for modification 10 April 1999/Accepted 25 May 1999

TB PNA FISH is a new fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for differentiation between species of the Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) in acid-fast bacillus-positive (AFB+) cultures is described. The test is based on fluorescein-labelled PNA probes that target the rRNA of MTC or NTM species applied to smears of AFB+ cultures for microscopic examination. Parallel testing with the two probes serves as an internal control for each sample such that a valid test result is based on one positive and one negative reaction. TB PNA FISH was evaluated with 30 AFB+ cultures from Denmark and 42 AFB+ cultures from Thailand. The MTC-specific PNA probe showed diagnostic sensitivities of 84 and 97%, respectively, and a diagnostic specificity of 100% in both studies, whereas the NTM-specific PNA probe showed diagnostic sensitivities of 91 and 64%, respectively, and a diagnostic specificity of 100% in both studies. The low sensitivity of the NTM-specific PNA probe in the Thai study was due to a relatively high prevalence of Mycobacterium fortuitum, which is not identified by the probe. In total, 63 (87%) of the cultures were correctly identified as MTC (n = 46) or NTM (n = 17), whereas the remaining 9 were negative with both probes and thus the results were inconclusive. None of the samples were incorrectly identified as MTC or NTM; thus, the predictive value of a valid test result obtained with TB PNA FISH was 100%.

* Corresponding author. Mailing address: Department of Microbiology, DAKO A/S, Produktionsvej 42, DK-2600 Glostrup, Denmark. Phone: 45 44 85 95 00. Fax: 45 44 92 00 56. E-mail: ole.feldballe{at}dako.dk.

Journal of Clinical Microbiology, September 1999, p. 2760-2765, Vol. 37, No. 9
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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