Identification of non-diphtheriae corynebacterium by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry.

We evaluated the Bruker Biotyper matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry for identification of 92 clinical isolates of Corynebacterium species in comparison to identification using rpoB or 16S rRNA gene sequencing. Eighty isolates (87%) yielded a score of ≥1.700, and all of these were correctly identified to the species level with the exception of Corynebacterium aurimucosum being misidentified as the closely related Corynebacterium minutissimum.

Comparison of direct colony method versus extraction method for identification of gram-positive cocci by use of Bruker Biotyper matrix-assisted laser desorption ionization-time of flight mass spectrometry.

We evaluated Bruker Biotyper (version 2.0) matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) for the identification of 305 clinical isolates of staphylococci, streptococci, and related genera by comparing direct colony testing with preparatory extraction. Isolates were previously identified by use of phenotypic testing and/or 16S rRNA gene sequencing. Manufacturer-specified score cutoffs for genus- and species-level identification were used. After excluding 7 isolates not present in the Biotyper library, the Biotyper correctly identified 284 (95%) and 207 (69%) isolates to the genus and species levels, respectively, using extraction. By using direct colony testing, the Biotyper identified 168 (56%) and 60 (20%) isolates to the genus and species levels, respectively. Overall, more isolates were identified to the genus and species levels with preparatory extraction than with direct colony testing (P < 0.0001). The analysis was repeated after dividing the isolates into two subgroups, staphylococci, streptococci, and enterococci (n = 217) and "related genera" (n = 81). For the former subgroup, the extraction method resulted in the identification of 213 (98%) and 171 (79%) isolates to the genus and species levels, respectively, whereas the direct colony method identified 136 (63%) and 56 (26%) isolates to the genus and species levels, respectively. In contrast, for the subgroup of related genera, the extraction method identified 71 (88%) and 36 (44%) isolates to the genus and species levels, respectively, while the direct colony method identified 32 (40%) and 4 (5%) isolates to the genus and species levels, respectively. For both subgroups, preparatory extraction was superior to direct colony testing for the identification of isolates to the genus and species levels (P < 0.0001). Preparatory extraction is needed for the identification of a substantial proportion of Gram-positive cocci using the Biotyper method according to manufacturer-specified score cutoffs.

VceR negatively regulates the vceCAB MDR efflux operon and positively regulates its own synthesis in Vibrio cholerae 569B.

The vceCAB (vce) operon encodes the multidrug resistance pump VceCAB (VCE), which contributes to resistance of Vibrio cholerae to carbonyl cyanide m-chlorophenylhydrazine (CCCP), deoxycholate, and pentachlorophenol by several-fold. vceR, which encodes the TetR-type repressor VceR and is divergently transcribed from vce, has been characterized in Escherichia coli. Detailed characterization of vceR in V. cholerae 569B confirmed the repressive effect of VceR on VCE function and indicated several novel features of VceR. Deletion of vceR increased resistance of strain 569B to CCCP and deoxycholate modestly, but did not affect resistance to pentachlorophenol. Transcriptional analysis revealed that vce expression was not only increased in strain 569BDeltavceR::Omega by 2-fold but continued to rise throughout the growth cycle. Using a vceR-lux transcriptional fusion plasmid, we examined whether vceR is autoregulated in strain 569B. Expression of vceR from the vceR-lux fusion was significantly lower in strain 569BDeltavceR::Omega than in strain 569B. In addition, exposure to CCCP reduced vceR expression from the vceR-lux fusion in strain 569B but not in strain 569BDeltavceR::Omega. Despite differences in the VceR binding site in strain 569B from the previously recognized 28 bp sequence in V. cholerae CVD101, purified recombinant VceR bound to the 24 bp sequence from strain 569B. We propose that VceR modulates vce expression by binding in vivo to the 24 bp sequence within the vceR-vce intergenic region; unlike many TetR repressors that are negatively autoregulated, VceR positively regulates vceR expression in trans.