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Journal of Clinical Microbiology, December 2001, p. 4477-4482, Vol. 39, No. 12
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.12.4477-4482.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Rapid Identification of Mycobacteria to the Species Level Using INNO-LiPA Mycobacteria, a Reverse Hybridization Assay

P. N. Suffys,1,* A. da Silva Rocha,1 M. de Oliveira,1 C. E. Dias Campos,2 A. M. Werneck Barreto,2 F. Portaels,3 L. Rigouts,3 G. Wouters,4 G. Jannes,4 G. van Reybroeck,4 W. Mijs,4 and B. Vanderborght4,5

Biochemistry and Molecular Biology Department, Oswaldo Cruz Institute, Oswaldo Cruz Foundation,1 Reference Center Professor Hélio Fraga,2 and Laboratory of Molecular Biology, HUCFF, Federal University of Rio de Janeiro,5 Rio de Janeiro, Brazil, and Institute of Tropical Medicine, Antwerp,3 and Innogenetics, Zwijnaarde,4 Belgium

Received 25 May 2001/Returned for modification 16 July 2001/Accepted 13 September 2001

INNO-LiPA Mycobacteria (LiPA; Innogenetics, Zwijnaarde, Belgium) is a kit for the simultaneous detection and identification of Mycobacterium species in culture and identifies the Mycobacterium tuberculosis complex, the M. avium complex (MAC), and the following Mycobacterium species: M. kansasii, M. avium, M. intracellulare, M. scrofulaceum, M. gordonae, M. xenopi, and the M. chelonae-M. abscessus complex. The assay, which targets the 16S-23S rRNA spacer region, was evaluated on 157 mycobacterial strains that had been identified by conventional techniques and PCR-restriction enzyme analysis of the hsp65 gene (PRA). Forty-seven reference strains consisting of 37 different species and 110 human clinical isolates were submitted to the test, and all were hybridized with the Mycobacterium genus probe (MYC) on the LiPA strip (100% sensitivity). Ninety-four isolates hybridized to their corresponding species- or complex-specific probes; only one isolate phenotypically identified as M. gordonae did not react with its specific probe (99.4% accuracy). Thirty-seven MAC strains were phenotypically identified to the complex level and to the species level by LiPA as M. avium (n = 18) or M. intracellulare (n = 7) or as belonging to the M. avium-M. intracellulare-M. scrofulaceum complex (n = 12). Of the last 12 strains, 10 had M. avium PRA patterns and 2 had M. intracellulare PRA patterns. Three isolates that had been identified as a single species by conventional identification were proven to be mixed cultures by the LiPA assay. The whole procedure can be performed in 1 working day, starting with the supernatant of a small amount of bacterial mass that had been treated by freezing and then boiling.

* Corresponding author. Mailing address: Laboratory of Molecular and Diagnosis of Infectious Diseases, DBBM, IOC, Fiocruz, Av. Brasil 4365, Manguinhos 21045-900, Rio de Janeiro, Brazil. Phone: 55-21-5984289. Fax: 55-21-2709997. E-mail: psuffys{at}ioc.fiocruz.br.

Journal of Clinical Microbiology, December 2001, p. 4477-4482, Vol. 39, No. 12
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.12.4477-4482.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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