Identification of Gallibacterium species by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry evaluated by multilocus sequence analysis

被引:41
作者
Alispahic, Merima [1 ]
Christensen, Henrik [2 ]
Hess, Claudia [1 ]
Razzazi-Fazeli, Ebrahim [3 ]
Bisgaard, Magne [2 ]
Hess, Michael [1 ]
机构
[1] Univ Vet Med, Clin Avian Reptile & Fish Med, Dept Farm Anim & Vet Publ Hlth, A-1210 Vienna, Austria
[2] Univ Copenhagen, Dept Vet Dis Biol, Fac Life Sci, DK-1168 Copenhagen, Denmark
[3] Univ Vet Med, VetOMICS Core Facil Res Prote & Metabol, A-1210 Vienna, Austria
关键词
Gallibacterium; MALDI-TOF MS; MLSA; Characterization; DESORPTION IONIZATION-TIME; 16S RIBOSOMAL-RNA; PASTEURELLA-HAEMOLYTICA; SP-NOV; FAMILY PASTEURELLACEAE; PHYLOGENETIC ANALYSIS; RESPIRATORY-TRACT; GEN.-NOV; CLASSIFICATION; ANATIS;
D O I
10.1016/j.ijmm.2011.03.001
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) whole-cell fingerprinting was used for characterization of 66 reference strains of Gallibacterium. The 4 recognised Gallibacterium species and Gallibacterium genomospecies 1 yielded reproducible and unique mass spectrum profiles, which were confirmed with Bruker Biotyper reference database version 3. The reproducibility of MALDI-TOF MS results were evaluated varying the age and storage of the cultures investigated. Reliable species identification was possible for up to 8 days of storage at 4 degrees C and less reliable if the bacteria were stored at room temperature (20 degrees C). However, if the strains were grown longer than 48 h at 37 degrees C under microaerobic atmosphere, poor identification results were obtained, due to changes in protein profile. The MALDI-TOF MS results of all 66 strains demonstrated 87.9% concordance with results based upon biochemical/physiological characterization. In addition, diversities outlined by MALDI-TOF MS were verified by sequencing the rpoB (n= 43), 16S rRNA (n =28), infB (n = 14), and recN (n=14) genes (multilocus sequence analysis, MLSA). In addition, discrepancies were observed between some of the genes sequenced. Results obtained demonstrated that MALDI-TOF MS fingerprinting represents a fast and reliable method for identification and differentiation of the 4 recognised Gallibacterium species and possible a fifth species Gallibacterium genomospecies 1, with applications in clinical diagnostics. (C) 2011 Elsevier GmbH. All rights reserved.
引用
收藏
页码:513 / 522
页数:10
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