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Differential phenotyping of Brucella species using a newly developed semi-automated metabolic system.

Al Dahouk S, Scholz HC, Tomaso H, Bahn P, Göllner C, Karges W, Appel B, Hensel A, Neubauer H, Nöckler K - BMC Microbiol. (2010)

Bottom Line: The comprehensive testing of metabolic activity allows cluster analysis within the genus Brucella.The biotyping system developed for the identification of Brucella 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Federal Institute for Risk Assessment, Diedersdorfer Weg 1, D-12277 Berlin, Germany. Sascha.Al-Dahouk@gmx.de

ABSTRACT

Background: 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 Brucella and their differentiation into species and biovars.

Results: 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 Brucella and a collection of 60 field isolates. Based on specific and stable reactions a 96-well "Brucella identification and typing" plate (Micronaut™) was designed and re-tested in 113 Brucella isolates and a couple of closely related bacteria.Brucella species and biovars revealed characteristic metabolic profiles and each strain showed an individual pattern. Due to their typical metabolic profiles a differentiation of Brucella isolates to the species level could be achieved. The separation of B. canis from B. suis bv 3, however, failed. At the biovar level, B. abortus bv 4, 5, 7 and B. suis bv 1-5 could be discriminated with a specificity of 100%. B. melitensis isolates clustered in a very homogenous group and could not be resolved according to their assigned biovars.

Conclusions: The comprehensive testing of metabolic activity allows cluster analysis within the genus Brucella. The biotyping system developed for the identification of Brucella 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.

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The Brucella specific Micronaut™ microtiter plate. Design of the newly developed Brucella specific Micronaut™ microtiter plate including 93 selected substances.
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Figure 2: The Brucella specific Micronaut™ microtiter plate. Design of the newly developed Brucella specific Micronaut™ microtiter plate including 93 selected substances.

Mentions: The comprehensive biotyping of the reference strains resulted in clusters agreeing in principle with the present conception of the genus Brucella (Figure 1A). A subset of 93 substances which preserved the clustering of the reference strains and achieved a satisfying discrimination was consecutively selected (Figures 2 and 1B). The newly configured 96-well plate assay tested for 29 aminopeptidases, 2 phosphatases, 4 glucosidases, 1 esterase, and the metabolism of 11 monosaccharides, 3 disaccharides, 7 sugar derivates, 15 amino acids, 11 organic acids, 1 salt, 1 amino acid derivate, 1 peptide, and 1 base. In addition, 6 classical reactions, i.e. nitrite, nitrate, pyrazinamidase, Voges-Proskauer medium, urease and H2S production, and three controls, i.e. peptidase control, pyrazinamidase control and assimilation control were included.


Differential phenotyping of Brucella species using a newly developed semi-automated metabolic system.

Al Dahouk S, Scholz HC, Tomaso H, Bahn P, Göllner C, Karges W, Appel B, Hensel A, Neubauer H, Nöckler K - BMC Microbiol. (2010)

The Brucella specific Micronaut™ microtiter plate. Design of the newly developed Brucella specific Micronaut™ microtiter plate including 93 selected substances.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2984481&req=5

Figure 2: The Brucella specific Micronaut™ microtiter plate. Design of the newly developed Brucella specific Micronaut™ microtiter plate including 93 selected substances.
Mentions: The comprehensive biotyping of the reference strains resulted in clusters agreeing in principle with the present conception of the genus Brucella (Figure 1A). A subset of 93 substances which preserved the clustering of the reference strains and achieved a satisfying discrimination was consecutively selected (Figures 2 and 1B). The newly configured 96-well plate assay tested for 29 aminopeptidases, 2 phosphatases, 4 glucosidases, 1 esterase, and the metabolism of 11 monosaccharides, 3 disaccharides, 7 sugar derivates, 15 amino acids, 11 organic acids, 1 salt, 1 amino acid derivate, 1 peptide, and 1 base. In addition, 6 classical reactions, i.e. nitrite, nitrate, pyrazinamidase, Voges-Proskauer medium, urease and H2S production, and three controls, i.e. peptidase control, pyrazinamidase control and assimilation control were included.

Bottom Line: The comprehensive testing of metabolic activity allows cluster analysis within the genus Brucella.The biotyping system developed for the identification of Brucella 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Federal Institute for Risk Assessment, Diedersdorfer Weg 1, D-12277 Berlin, Germany. Sascha.Al-Dahouk@gmx.de

ABSTRACT

Background: 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 Brucella and their differentiation into species and biovars.

Results: 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 Brucella and a collection of 60 field isolates. Based on specific and stable reactions a 96-well "Brucella identification and typing" plate (Micronaut™) was designed and re-tested in 113 Brucella isolates and a couple of closely related bacteria.Brucella species and biovars revealed characteristic metabolic profiles and each strain showed an individual pattern. Due to their typical metabolic profiles a differentiation of Brucella isolates to the species level could be achieved. The separation of B. canis from B. suis bv 3, however, failed. At the biovar level, B. abortus bv 4, 5, 7 and B. suis bv 1-5 could be discriminated with a specificity of 100%. B. melitensis isolates clustered in a very homogenous group and could not be resolved according to their assigned biovars.

Conclusions: The comprehensive testing of metabolic activity allows cluster analysis within the genus Brucella. The biotyping system developed for the identification of Brucella 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.

Show MeSH
Related in: MedlinePlus