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Detection and identification of Mycobacterium species isolates by DNA microarray
Masao Fukushima1, Kenichi Kakinuma, Hiroshi Hayashi
1Genomics Research Institute, SRL, Inc., 5-6-50 Shinmachi, Hino-shi, Tokyo 191-0002, Japan.
Journal of Clinical Microbiology
|June 7, 2003
Summary
A new DNA gyrase B subunit (gyrB) gene microarray rapidly identifies Mycobacterium species. This method aids in the effective diagnosis and management of tuberculosis by distinguishing closely related bacteria.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Rapid identification of Mycobacterium species is crucial for effective tuberculosis management.
- Analysis of the DNA gyrase B subunit (gyrB) gene offers a promising approach for bacterial species identification.
Purpose of the Study:
- To develop and evaluate a microarray assay utilizing gyrB gene sequences for the identification of Mycobacterium species.
- To assess the assay's ability to differentiate closely related species and its concordance with conventional methods.
Main Methods:
- PCR amplification of a conserved gyrB gene region from clinical samples using specific primers.
- Development of oligonucleotide probes targeting specific gyrB gene regions for 14 Mycobacterium species.
- Hybridization of fluorescently labeled amplified DNA with probes spotted on a microarray slide.
Main Results:
- The gyrB-based microarray generated unique hybridization patterns for each tested Mycobacterium species.
- The assay successfully differentiated closely related mycobacterial species.
- Results showed good correlation with conventional culture methods for mycobacteria detection.
Conclusions:
- A gyrB-based microarray provides a rapid and accurate method for identifying Mycobacterium species, including closely related ones.
- This diagnostic tool holds potential for improving the management of tuberculosis.