Uncommonly isolated clinical <Emphasis Type="Italic">Pseudomonas</Emphasis>: identification and phylogenetic assignation |
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Authors: | M Mulet M Gomila A Ramírez S Cardew E R B Moore J Lalucat E García-Valdés |
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Institution: | 1.Department of Biology, Microbiology, Edificio Guillem Colom,Universitat de les Illes Balears,Palma de Mallorca,Spain;2.Servicio de Microbiología,Hospital Universitario Son Espases,Palma de Mallorca,Spain;3.Department of Infectious Disease, Institute of Biomedicine,Sahlgrenska Academy of the University of Gothenburg,Gothenburg,Sweden;4.Institut Mediterrani d’Estudis Avan?ats (IMEDEA, CSIC-UIB),Palma de Mallorca,Spain |
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Abstract: | Fifty-two Pseudomonas strains that were difficult to identify at the species level in the phenotypic routine characterizations employed by clinical microbiology laboratories were selected for genotypic-based analysis. Species level identifications were done initially by partial sequencing of the DNA dependent RNA polymerase sub-unit D gene (rpoD). Two other gene sequences, for the small sub-unit ribosonal RNA (16S rRNA) and for DNA gyrase sub-unit B (gyrB) were added in a multilocus sequence analysis (MLSA) study to confirm the species identifications. These sequences were analyzed with a collection of reference sequences from the type strains of 161 Pseudomonas species within an in-house multi-locus sequence analysis database. Whole-cell matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analyses of these strains complemented the DNA sequenced-based phylogenetic analyses and were observed to be in accordance with the results of the sequence data. Twenty-three out of 52 strains were assigned to 12 recognized species not commonly detected in clinical specimens and 29 (56 %) were considered representatives of at least ten putative new species. Most strains were distributed within the P. fluorescens and P. aeruginosa lineages. The value of rpoD sequences in species-level identifications for Pseudomonas is emphasized. The correct species identifications of clinical strains is essential for establishing the intrinsic antibiotic resistance patterns and improved treatment plans. |
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