TY - JOUR
T1 - Identification of Gallibacterium speciesby matrix-assisted laser desorption/ ionization time-of-flight mass spectrometryevaluated bymultilocus sequence analysis
AU - Alispahic, Merima
AU - Christensen, Henrik
AU - Hess, Claudia
AU - Razzazi-Fazeli, Ebrahim
AU - Bisgaard, Magne
AU - Hess, Michael
PY - 2011/8
Y1 - 2011/8
N2 - 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 °C and less reliable if the bacteria were stored at room temperature (20 °C). However, if the strains were grown longer than 48. h at 37 °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.
AB - 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 °C and less reliable if the bacteria were stored at room temperature (20 °C). However, if the strains were grown longer than 48. h at 37 °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.
KW - Former LIFE faculty
KW - Gallibacterium
KW - MALDI-TOF MS
KW - MLSA
KW - Characterization
U2 - 10.1016/j.ijmm.2011.03.001
DO - 10.1016/j.ijmm.2011.03.001
M3 - Journal article
SN - 1438-4221
VL - 301
SP - 513
EP - 522
JO - International Journal of Medical Microbiology
JF - International Journal of Medical Microbiology
IS - 6
ER -