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Published online ahead of print on 22 October 2009 as doi:10.1099/mic.0.030684-0
Microbiology (2009), DOI 10.1099/mic.0.030684-0
© 2009 Society for General Microbiology
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Microbiology 0 (2009), mic.0.030684; DOI  10.1099/mic.0.030684-0
© 2009 Society for General Microbiology

Molecular Typing of Mycobacterium intracellulare using Multilocus Variable-Number of Tandem-Repeat Analysis: Identification of Loci and Analysis of Clinical Isolates

Kazuya Ichikawa1, Tetsuya Yagi2, Takayuki Inagaki3, Makoto Moriyama3, Taku Nakagawa1, Kei-ichi Uchiya3, Toshiaki Nikai3 and Kenji Ogawa4,5

1 National Hospital Organization, Higashinagoya National Hospital;
2 Nagoya University Hospital,;
3 Meijo University;
4 National Hospital Organization, Higashinagoya National Hospital,

In addition to its known status as a disseminated disease in HIV-positive patients, Mycobacterium avium complex (MAC) is increasingly recognized as a causative pathogen of respiratory disease in HIV-negative patients. MAC is divided into Mycobacterium avium, and the less-epidemiologically studied Mycobacterium intracellulare. Genetic typing for M. intracellulare using variable number of tandem repeats (VNTR) has not yet been developed. The aim of this study was to identify VNTR loci in the genome of Mycobacterium intracellulare and apply them as an epidemiological tool to clinical isolates. Here, we identified 25 VNTR loci on the Mycobacterium intracellulare genome, of which 16 showed variations among clinical isolates in the number of tandem repeat motifs. Among the 74 Mycobacterium intracellulare isolates, 50 genotypes were distinguished using the 16 VNTR loci, resulting in a Hunter Gaston's Discriminatory Index of 0.988. Moreover, all 16 VNTR loci were stable in different sets of isolates recovered within time intervals ranging from 2 to 1551 days from 14 separate patients. These results indicate that for use as epidemiological markers of Mycobacterium intracellulare, the loci in this VNTR assay are highly discriminating and stable over time.

5 E-mail: ogawak{at}toumei.hosp.go.jp






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