Caracterización molecular de aislamientos de enterobacter cloacae multirresistentes, productores ß-Lactamasas provenientes de pacientes de un hospital de tercer nivel de Bogotá
Molecular characterizacion of multi-cephalosporin resistan Enterobacter cloacae isolates from a third level hospital in Bogota-Colombia
Palabras clave:
enterobacter cloacae, cefalosporinas, ß-lactamasas, isoeléctricos (Mezclas anfolíticas), clones (células clonares), infección nosocomial (infección hospitalaria). (es)enterobacter cloacae, cephalosporins, beta-lactamases, ampholyte mixtures, clone cells, infection, cross infection. (en)
Antecedentes. Las enterobacterias, antaño flora normal del tracto gastrointestinal, han cambiado su biología y emergido como agentes patógenos nosocomiales que se tornan resistentes los antibióticos conocidos.
Objetivo. Realizar la caracterización epidemiológico-molecular de 20 aislamientos de Enterobacter cloacae resistentes a cefalosporinas de tercera generación; provenientes de un hospital de tercer nivel de Bogotá-Colombia.
Material y métodos. Los aislamientos fueron identificados mediante sistemas automatizados Microscan y VITEK, se utilizó el Enterobacter asbureae como control externo inter-especie.
La confirmación de resistencia se hizo por técnica de difusión en agar, y una vez establecida se realizó BLEE para comprobación. La determinación de puntos isoeléctricos se hizo, mediante lisis por ultrasonido y la genotipificación mediante la metodología para bacterias Gramnegativas propuesta por Versalovic.
Resultados: Los aislamientos colectados durante un año fueron causantes de 15 casos de infección Intrahospitalaria y dos colonizaciones.
Todos los aislamientos presentaron resistencia a cefotaxima, ceftazidima, ceftriaxona, aztreonam y ciprofloxacina, 95% a amikacina, gentamicina y cloranfenicol, 75% a trimetoprim/sulfametoxazol, 20% a cefepime y todos fueron sensibles a imipenem.
Dos aislamientos fueron confirmados como productores de â-lactamasas de espectro extendido (BLEE) por la técnica microbiológica de disco combinado.
Por isoelectroenfoque presentaron dos â-lactamasas con puntos isoeléctricos (pI) de 5,4 y 8,2. En los 18 aislamientos no inhibidos por ácido clavulánico, se detectaron entre 2 y 4 â-lactamasas con pI de 5,4; 6,0; 7,0; 8,2 y mayor que 8,2; la resistencia a cefalosporinas de tercera generación podría ser atribuida a la hiperproducción de AmpC; los valores de pI sugieren la producción simultánea de â-lactamasas tipo SHV y TEM. La genotipificación mediante tres metodologías de rep-PCR (ERIC; REP y BOX) agrupó la población estudiada en siete clones: seis constituidos por un solo aislamiento y el clon predominante E1/B1/R1 agrupó 14 aislamientos causantes de infección en diez pacientes.
Conclusión. Se identificó un clon de Enterobacter cloacae multirresistente, endémico en una institución de tercer nivel en Bogotá, causante de infección nosocomial y quirúrgica en particular.
Background. Enterobacter species were normal in gastrointestinal tract, but nowadays, its biology has changed and there are nosocomial agents with antibiotics resistance.
Objective. To make an epidemiological and molecular characterization of 20 isolates of Enterobacter cloacae with third generation cephalosporin resistance, from a hospital of third level in Bogotá-Colombia.
Material and methods. Isolates were identified with Microscan and VITEK, Enterobacter asbureae was utilized as an inter-specie control.
Resistance was confirmed by agar diffusion and by BLEE techniques. Isoelectric points were determined by ultrasound lyses and genotypication by Versalovic´s system for gram negative bacteria.
Results. The isolates collected over the course of a year caused 15 cases of intra-hospital infection and two colonisations. All isolates presented resistance to cefotaxime, ceftazidime, ceftriaxone, aztreonam and ciprofloxacin, 95% to amikacin, gentamicin and chloramphenicol, 75% to trimethoprim/ sulphamethoxazole, 20% to cefepime and all were sensitive to imipenem. Two isolates were confirmed as extended spectre â-Iactamase (ESBL) producers by microbiologic al combined disktechnique; two â-Iactamases having 5.4 and 8.2 isoelectric points (pI) were presented by isoelectric focusing. Between 2 and 4 â-Iactamases having 5.4, 6.0, 7.0, 8.2 and >8.2 pl were detected in the 18 isolates which were not inhibited by clavulanic acid. Third-generation cephalosporin-resistance was attributed to AmpC hyper-production; pl values suggested simultaneous SHV and TEM â lactamase production. Genotyping by three rep- PCR methodologies (ERIC, REP and BOX) grouped the population studied into 7clones; 6 were constitute d by a single isolate and the predominant E1/B1/R1 clone grouped 14 isolates causing infection in 10 patients. This work led to a multiresistant Enterobacter cloacae clone being detected, considered endemic for the institution, in the studied surgical patients it was una predominantly cause of intrahospital infection.
Conclusion. We detected a clone of Enterobacter cloacae with multi-cephalosporin resistance, has an endemic strain in a hospital of third level in Bogotá, who caused nosocomial infection, in special of surgical patients.
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