Detection and Identification of Mycobacterium Species Isolates by DNA Microarray
- 1 June 2003
- journal article
- research article
- Published by American Society for Microbiology in Journal of Clinical Microbiology
- Vol. 41 (6), 2605-2615
- https://doi.org/10.1128/jcm.41.6.2605-2615.2003
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.Keywords
This publication has 29 references indexed in Scilit:
- Caloramator viterbensis sp. nov., a novel thermophilic, glycerol-fermenting bacterium isolated from a hot spring in ItalyInternational Journal of Systematic and Evolutionary Microbiology, 2002
- Evaluation of Dry and Wet Transported Intravaginal Swabs in Detection of Chlamydia trachomatis and Neisseria gonorrhoeae Infections in Female Soldiers by PCRJournal of Clinical Microbiology, 2002
- Diagnosis of foot-and-mouth disease by RT-PCR: use of phylogenetic data to evaluate primers for the typing of viral RNA in clinical samplesArchiv für die gesamte Virusforschung, 2001
- Direct Identification of Mycobacteria from MB/BacT Alert 3D Bottles: Comparative Evaluation of Two Commercial Probe AssaysJournal of Clinical Microbiology, 2001
- Performance Assessment of New Multiplex Probe Assay for Identification of MycobacteriaJournal of Clinical Microbiology, 2001
- Identification of Clinical isolates of Mycobacterium spp. by sequence analysis of the 16S ribosomal RNA geneAPMIS, 1999
- Phylogenetic relationships of Pseudomonas putida strains deduced from the nucleotide sequences of gyrB, rpoD and 16S rRNA genesInternational Journal of Systematic and Evolutionary Microbiology, 1998
- Phylogenetic Analysis of Acinetobacter Strains Based on the Nucleotide Sequences of gyrB Genes and on the Amino Acid Sequences of Their ProductsInternational Journal of Systematic and Evolutionary Microbiology, 1996
- Rapid diagnosis of tuberculous meningitis by polymerase chain reactionThe Lancet, 1991
- Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA PolymeraseScience, 1988