title: Detection and identification of Mycobacterium species isolates by DNA microarray.
abstract: 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.
authors.affiliation: Genomics Research Institute, SRL, Inc., 5-6-50 Shinmachi, Hino-shi, Tokyo 191-0002, Japan.
authors.forename: Masao
authors.initials: M
authors.lastname: Fukushima
authors.forename: Kenichi
authors.initials: K
authors.lastname: Kakinuma
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Hayashi
authors.forename: Hiroko
authors.initials: H
authors.lastname: Nagai
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Ito
authors.forename: Ryuji
authors.initials: R
authors.lastname: Kawaguchi
mesh_terms: D001483:Base Sequence; D003470:Culture Media; D027081:DNA Gyrase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D009164:Mycobacterium Infections / Q000382:microbiology; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes; D013045:Species Specificity
chemical_list: D003470:Culture Media; D015345:Oligonucleotide Probes; D027081:DNA Gyrase