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Journal of Clinical Microbiology, September 2001, p. 3222-3227, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3222-3227.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Direct Identification of Mycobacteria from MB/BacT Alert 3D Bottles: Comparative Evaluation of Two Commercial Probe Assays

Claudio Scarparo,1,* Paola Piccoli,1 Alessandra Rigon,1 Giuliana Ruggiero,1 Domenico Nista,2 and Claudio Piersimoni2

Regional Mycobacteria Reference Centre, Laboratory of Clinical Microbiology and Virology, San Bortolo Hospital, Vicenza,1 and Department of Clinical Microbiology, General Hospital Umberto I---Torrette, Ancona,2 Italy

Received 23 April 2001/Returned for modification 7 June 2001/Accepted 28 June 2001

The new INNO-LiPA Mycobacteria (Innogenetics, Ghent, Belgium), a reverse-hybridization-based line probe assay, and the AccuProbe assay (Gen-Probe Inc., San Diego, Calif.) were applied to MB/BacT Alert 3D (MB/BacT) system (Organon Teknika, Boxtel, The Netherlands) culture bottles and evaluated for mycobacterial identification. From 2,532 respiratory and extrapulmonary specimens submitted for culture, 168 were flagged positive by the MB/BacT system and promptly evaluated for identification (within 24 h). Each of 163 vials grew one mycobacterial isolate, including Mycobacterium tuberculosis complex (n = 73), M. avium complex (n = 3), M. avium (n = 8), M. intracellulare (n = 5), M. kansasii (n = 15), M. gordonae (n = 8), M. malmoense (n = 3), M. chelonae (n = 13), M. abscessus (n = 2), M. xenopi (n = 11), M. scrofulaceum (n = 2), M. fortuitum (n = 7), M. terrae (n = 3), M. simiae (n = 2), M. celatum (n = 3), M. flavescens (n = 1), M. interjectum (n = 1), M. bohemicum (n = 1), and M. pulveris (n = 2). Five cultures yielded mixed growth of two mycobacterial species: M. tuberculosis complex plus M. gordonae (n = 2), M. tuberculosis complex plus M. chelonae (n = 1), M. tuberculosis complex plus M. xenopi (n = 1), and M. avium plus M. chelonae (n = 1). In testing of one-isolate vials, both systems showed excellent sensitivity and specificity for all species and complexes for which they are licensed (nine for INNO-LiPA Mycobacteria versus six for AccuProbe). There were minor discrepancies in results for two isolates identified by INNO-LiPA Mycobacteria as M. avium - M. intracellulare - M. scrofulaceum (MAIS) complex and by AccuProbe as M. intracellulare. In testing of two-isolate vials, INNO-LiPA Mycobacteria correctly identified all isolates, while the AccuProbe assay failed to identify three M. tuberculosis complex isolates and one M. avium isolate. The AccuProbe assay was completed within 2 h, while INNO-LiPA Mycobacteria required a 6-h period. In our opinion, INNO-LiPA Mycobacteria offers the following advantages: (i) it contains a genus-specific probe that, in addition to being used in genus identification, may be used as an internal control for both the amplification and hybridization steps; (ii) it simultaneously identifies M. tuberculosis complex, MAIS complex, and seven other mycobacterial species, even from mixed cultures; (iii) its mycobacterial DNA amplification ensures reliable results independent from the concentration of viable microorganisms; and (iv) it genotypically identifies M. kansasii and M. chelonae. In conclusion, even though INNO-LiPA Mycobacteria is considerably less easy to use than AccuProbe, requiring personnel skilled in molecular biology techniques, it represents an excellent approach for routine identification of frequently encountered mycobacteria.

* Corresponding author. Mailing address: Regional Mycobacteria Reference Centre, Microbiology and Virology Laboratory, San Bortolo Hospital, Viale Rodolfi 37, Vicenza I-36100, Italy. Phone: 39 0444 993507. Fax: 39 0444 993963. E-mail: claudio.scarparo{at}tin.it.

Journal of Clinical Microbiology, September 2001, p. 3222-3227, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3222-3227.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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