RESUMO
La espectrometría de masas (EM) (matrix assisted laser desorption ionization-time of flight) MALDI-TOF demostró ser una herramienta robusta para la identificación de numerosos grupos taxonómicos. No obstante, presenta limitaciones. Una ventaja clave de la técnica es la flexibilidad para la incorporación de espectros proteicos de microorganismos ausentes en la base de datos comercial. Dada la prevalencia de Burkholderia contaminans en los pacientes fibroquísticos en Argentina, y a que en ellos es crucial el diagnóstico microbiológico rápido y confiable, la EM MALDI-TOF surge como una herramienta estratégica. El objetivo del trabajo fue desarrollar una base de datos adicional con espectros peptídicos de aislamientos de referencia de B. contaminans. La misma demostró ser exitosa para la identificación del 97% de los aislamientos analizados. Por lo cual la EM MALDI-TOF con la base de datos extendida resultó ser una herramienta útil para la identificación y diferenciación de otras especies relacionadas a B. contaminans.
MALDI-TOF (matrix assisted laser desorption ionization-time of flight) mass spectrometry (MS) proved to be a robust tool for the identification of numerous taxonomic groups. However, it has limitations. A key advantage of this technique is the flexibility for the incorporation of protein profiles of microorganisms not included in the commercial database. Due to the prevalence of Burkholderia contaminans in fibrocystic patients in Argentina and the fact that rapid and reliable microbiological diagnosis is crucial in them, MALDI-TOF MS emerges as a strategic tool. The aim of this work was to develop an additional database with peptide spectra of reference isolates of B. contaminans. This database demonstrated to be successful for the identification of 97% of the isolates analyzed. Therefore, MALDI-TOF MS with the extended database was a useful tool for the identification and differentiation of other related species to B. contaminans.
Assuntos
Humanos , Bases de Dados Factuais , Técnicas Bacteriológicas , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Burkholderia/isolamento & purificação , Especificidade da Espécie , Proteínas de Bactérias/análise , Algoritmos , Reprodutibilidade dos Testes , Infecções por Burkholderia/complicações , Infecções por Burkholderia/microbiologia , Burkholderia/classificação , Burkholderia/química , Fibrose Cística/complicações , Fibrose Cística/microbiologiaRESUMO
Fifteen strains of Burkholderia (Pseudomonas) solanacearum were isolated, identified from different parts of India and screened for plasmid profile. They harboured 1-3 plasmids, but most of the isolates contained a single plasmid. Plasmids were classified on the basis of replicon typing by Southern hybridization using incompatible specific rep probes. Plasmids of 9 strains hybridized with repP probe, plasmids of 5 strains hybridized with repW probe and one plasmid hybridized with repQ probe. Fingerprint and cluster analyses of plasmids from 10 strains of B. solanacearum digested with Eco RI revealed that plasmids of race 3 strains clustered in one group and the plasmids of race 1 strains clustered in another group.