Abstract
Summary The specificity of previously published oligonucleotide probes war re-evaluated based on the expanded 16S and 23S rRNA databases. Moreover, new genus- and (sub) species-specific 16S rRNA-targeted oligonucleotide probes were designed in a computer-assisted strategy using the PROBE DESIGN tool of the software package ARB. Probes for Pseudomonas stutzeri and for the genera Acidovorax, Comamonas and Hydrogenophaga were evaluated. Optimal hybridization conditions for the probes have been determined. Using these conditions the probes were tested for binding to 16S rRNA of reference cultures. Furthermore, fluorescently labeled derivatives of these probes were tested for their applicability for whole cell hybridization. These rRNA-targeted probes are the basis for high-resolution, rapid, automated microbial identification.
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