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Journal of Clinical Microbiology, June 2003, p. 2605-2615, Vol. 41, No. 6
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.6.2605-2615.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Detection and Identification of Mycobacterium Species Isolates by DNA Microarray

Masao Fukushima,1,2 Kenichi Kakinuma,1,2 Hiroshi Hayashi,1 Hiroko Nagai,3 Kunihiko Ito,4 and Ryuji Kawaguchi1*

Genomics Research Institute,1 Center for Molecular Biology and Cytogenetics,2 Laboratory of Infection and Immunology, SRL, Inc., 5-6-50 Shinmachi, Hino-shi, Tokyo 191-0002,3 Clinical Research Division, Research Institute of Tuberculosis, Department of Respiratory Disease, Fukujuji Hospital, Japan Anti-Tuberculosis Association, 3-1-24 Matsuyama, Kiyose-shi, Tokyo 204-8533, Japan4

Received 12 July 2002/ Returned for modification 3 October 2002/ Accepted 13 February 2003

Rapid identification of Mycobacterium species isolates is necessary for the effective management of tuberculosis. Recently, analysis of DNA gyrase B subunit (gyrB) genes has been identified as a suitable means for the identification of bacterial species. We describe a microarray assay based on gyrB gene sequences that can be used for the identification of Mycobacteria species. Primers specific for a gyrB gene region common to all mycobacteria were synthesized and used for PCR amplification of DNA purified from clinical samples. A set of oligonucleotide probes for specific gyrB gene regions was developed for the identification of 14 Mycobacterium species. Each probe was spotted onto a silylated glass slide with an arrayer and used for hybridization with fluorescently labeled RNA derived from amplified sample DNA to yield a pattern of positive spots. This microarray produced unique hybridization patterns for each species of mycobacteria and could differentiate closely related bacterial species. Moreover, the results corresponded well with those obtained by the conventional culture method for the detection of mycobacteria. We conclude that a gyrB-based microarray can rapidly detect and identify closely related mycobacterial species and may be useful in the diagnosis and effective management of tuberculosis.

* Corresponding author. Mailing address: Genomics Research Institute, SRL, Inc., 5-6-50 Shinmachi, Hino-shi, Tokyo 191-0002, Japan. Phone: 81-426-48-3873. Fax: 81-426-48-4054. E-mail: ryukaw{at}srl.srl-inc.co.jp.

Journal of Clinical Microbiology, June 2003, p. 2605-2615, Vol. 41, No. 6
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.6.2605-2615.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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