<p>Abstract</p> <p>Background</p> <p>A commercial biotyping system (Taxa Profile™, Merlin Diagnostika) testing the metabolization of various substrates by bacteria was used to determine if a set of phenotypic features will allow the identification of members of the genus <it>Brucella </it>and their differentiation into species and biovars.</p> <p>Results</p> <p>A total of 191 different amines, amides, amino acids, other organic acids and heterocyclic and aromatic substrates (Taxa Profile™ A), 191 different mono-, di-, tri- and polysaccharides and sugar derivates (Taxa Profile™ C) and 95 amino peptidase- and protease-reactions, 76 glycosidase-, phosphatase- and other esterase-reactions, and 17 classic reactions (Taxa Profile™ E) were tested with the 23 reference strains representing the currently known species and biovars of <it>Brucella </it>and a collection of 60 field isolates. Based on specific and stable reactions a 96-well "<it>Brucella </it>identification and typing" plate (Micronaut™) was designed and re-tested in 113 <it>Brucella </it>isolates and a couple of closely related bacteria.</p> <p><it>Brucella </it>species and biovars revealed characteristic metabolic profiles and each strain showed an individual pattern. Due to their typical metabolic profiles a differentiation of <it>Brucella </it>isolates to the species level could be achieved. The separation of <it>B. canis </it>from <it>B. suis </it>bv 3, however, failed. At the biovar level, <it>B. abortus </it>bv 4, 5, 7 and <it>B. suis </it>bv 1-5 could be discriminated with a specificity of 100%. <it>B. melitensis </it>isolates clustered in a very homogenous group and could not be resolved according to their assigned biovars.</p> <p>Conclusions</p> <p>The comprehensive testing of metabolic activity allows cluster analysis within the genus <it>Brucella</it>. The biotyping system developed for the identification of <it>Brucella </it>and differentiation of its species and biovars may replace or at least complement time-consuming tube testing especially in case of atypical strains. An easy to handle identification software facilitates the applicability of the Micronaut™ system for microbiology laboratories.</p
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