Lista, Florigio;
Reubsaet, Frans AG;
De Santis, Riccardo;
Parchen, Rene R;
de Jong, Ad L;
Kieboom, Jasper;
van der Laaken, Anton L;
Voskamp-Visser, Ingrid AI;
Fillo, Silvia;
Jansen, Hugo-Jan;
Van der Plas, Jan;
Paauw, Armand
Reliable identification at the species level of Brucella isolates with MALDI-TOF-MS
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Medientyp:
E-Artikel
Titel:
Reliable identification at the species level of Brucella isolates with MALDI-TOF-MS
Beteiligte:
Lista, Florigio;
Reubsaet, Frans AG;
De Santis, Riccardo;
Parchen, Rene R;
de Jong, Ad L;
Kieboom, Jasper;
van der Laaken, Anton L;
Voskamp-Visser, Ingrid AI;
Fillo, Silvia;
Jansen, Hugo-Jan;
Van der Plas, Jan;
Paauw, Armand
Erschienen:
Springer Science and Business Media LLC, 2011
Beschreibung:
<jats:title>Abstract</jats:title>
<jats:sec>
<jats:title>Background</jats:title>
<jats:p>The genus <jats:italic>Brucella</jats:italic> contains highly infectious species that are classified as biological threat agents. The timely detection and identification of the microorganism involved is essential for an effective response not only to biological warfare attacks but also to natural outbreaks. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is a rapid method for the analysis of biological samples. The advantages of this method, compared to conventional techniques, are rapidity, cost-effectiveness, accuracy and suitability for the high-throughput identification of bacteria. Discrepancies between taxonomy and genetic relatedness on the species and biovar level complicate the development of detection and identification assays.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Results</jats:title>
<jats:p>In this study, the accurate identification of <jats:italic>Brucella</jats:italic> species using MALDI-TOF-MS was achieved by constructing a <jats:italic>Brucella</jats:italic> reference library based on multilocus variable-number tandem repeat analysis (MLVA) data. By comparing MS-spectra from <jats:italic>Brucella</jats:italic> species against a custom-made MALDI-TOF-MS reference library, MALDI-TOF-MS could be used as a rapid identification method for <jats:italic>Brucella</jats:italic> species. In this way, 99.3% of the 152 isolates tested were identified at the species level, and <jats:italic>B. suis</jats:italic> biovar 1 and 2 were identified at the level of their biovar. This result demonstrates that for <jats:italic>Brucella</jats:italic>, even minimal genomic differences between these serovars translate to specific proteomic differences.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Conclusions</jats:title>
<jats:p>MALDI-TOF-MS can be developed into a fast and reliable identification method for genetically highly related species when potential taxonomic and genetic inconsistencies are taken into consideration during the generation of the reference library.</jats:p>
</jats:sec>