Oligonucleotide microarray for identification of Bacillus anthracis based on intergenic transcribed spacers in ribosomal DNA

We developed a DNA microarray for identification of Bacillus anthracis and other phylogenetic groupings within the Bacillus cereus group. Nucleotide sequences of 16S-235 ribosomal DNA internal transcribed spacers containing genes for tRNA(I1e) from 52 B. anthracis strains were found to be identical to sequences from seven strains published previously and different from all other bacteria. When 42 oligonucleotide probes targeting polymorphic sites were immobilized on glass slides and hybridized to fluorescently labeled PCR amplification products, one or more mismatches could be discriminated in all but one cases. Hence, hybridization events were highly specific and identification of B. anthracis was straightforward. (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.