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

doi:10.1128/JCM.39.9.3222-3227.2001

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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 AccuProbeassay (Gen-Probe Inc., San Diego, Calif.) were applied to MB/BacTAlert 3D (MB/BacT) system (Organon Teknika, Boxtel, The Netherlands)culture bottles and evaluated for mycobacterial identification.From 2,532 respiratory and extrapulmonary specimens submittedfor culture, 168 were flagged positive by the MB/BacT system andpromptly evaluated for identification (within 24 h). Each of 163vials grew one mycobacterial isolate, including Mycobacteriumtuberculosis 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 mixedgrowth of two mycobacterial species: M. tuberculosis complex plusM. 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 forall species and complexes for which they are licensed (nine forINNO-LiPA Mycobacteria versus six for AccuProbe). There were minordiscrepancies in results for two isolates identified by INNO-LiPAMycobacteria as M. avium - M. intracellulare - M. scrofulaceum(MAIS) complex and by AccuProbe as M. intracellulare. In testingof two-isolate vials, INNO-LiPA Mycobacteria correctly identifiedall isolates, while the AccuProbe assay failed to identify threeM. tuberculosis complex isolates and one M. avium isolate. TheAccuProbe assay was completed within 2 h, while INNO-LiPA Mycobacteriarequired a 6-h period. In our opinion, INNO-LiPA Mycobacteriaoffers the following advantages: (i) it contains a genus-specificprobe that, in addition to being used in genus identification,may be used as an internal control for both the amplificationand hybridization steps; (ii) it simultaneously identifies M.tuberculosis complex, MAIS complex, and seven other mycobacterialspecies, even from mixed cultures; (iii) its mycobacterial DNAamplification ensures reliable results independent from the concentrationof viable microorganisms; and (iv) it genotypically identifiesM. kansasii and M. chelonae. In conclusion, even though INNO-LiPAMycobacteria is considerably less easy to use than AccuProbe,requiring personnel skilled in molecular biology techniques, itrepresents an excellent approach for routine identification offrequently encounteredmycobacteria.

^* Corresponding author. Mailing address: Regional Mycobacteria Reference Centre, Microbiology and Virology Laboratory, San BortoloHospital, Viale Rodolfi 37, Vicenza I-36100, Italy. Phone: 390444 993507. Fax: 39 0444 993963. E-mail: claudio.scarparo{at}tin.it.

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