RESUMO
OBJECTIVE: To typify by molecular and phenotypical methods, MRSA strains, isolated from patients and nurses to establish their possible clonal origin. MATERIALS AND METHODS: 50 MRSA strains isolated in a teaching hospital in Maracaibo (Venezuela) were analyzed. The typification of MRSA strains was performed by means of pulsed-field gel electrophoresis (PFGE) and antibiotyping. RESULTS: In patients, 12 clusters (I-XII) and 19 antibiotypes were found; whereas in the health-care personnel, 6 clusters (I-VI) and two antbiotypes were detected. There was no statistically significative association between antibiotypes and band patterns obtained by PFGE (p>0.05). CONCLUSIONS: By means of detection of resistance markers and PFGE, it is feasible to discrimínate the nature of the clinical strains of MRSA. The obtained results show the possible nosocomial transmission of MRSA strains and their clonal spread in hospital departments, particularly at the ICU.
Assuntos
DNA Bacteriano/análise , Staphylococcus aureus Resistente à Meticilina/genética , Antibacterianos/farmacologia , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Eletroforese em Gel de Campo Pulsado , Genótipo , Hospitais Universitários , Humanos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Recursos Humanos em Hospital , FenótipoRESUMO
Objective: To typify by molecular and phenotypical methods, MRSA strains, isolated from patients and nurses to establish their possible clonal origin. Materials and Methods: 50 MRSA strains isolated in a teaching hospital in Maracaibo (Venezuela) were analyzed. The typification of MRSA strains was performed by means of pulsed-field gel electrophoresis (PFGE) and antibiotyping. Results: In patients, 12 clusters (I-XII) and 19 antibiotypes were found; whereas in the health-care personnel, 6 clusters (I-VI) and two antbiotypes were detected. There was no statistically significative association between antibiotypes and band patterns obtained by PFGE (p>0.05). Conclusions: By means of detection of resistance markers and PFGE, it is feasible to discrimínate the nature of the clinical strains of MRSA. The obtained results show the possible nosocomial transmission of MRSA strains and their clonal spread in hospital departments, particularly at the ICU.
Objetivo: Tipificar por métodos moleculares y fenotípicos, las cepas SAMR aisladas de pacientes y personal de enfermería para establecer su posible origen clonal. Materiales y Métodos: Se analizaron 50 cepas SAMR aisladas en un Hospital Universitario de Maracaibo (Venezuela). La tipificación se efectuó mediante electroforesis en gel de campo pulsado (EGCP) y antibiotipia. Resultados: Entre pacientes, se obtuvieron 19 antibiotipos y 12 grupos (I-XII); mientras que, en el personal de salud, por EGCP se detectaron seis grupos (I-VI) y dos antibiotipos. No se encontró asociación estadísticamente significativa entre los antibiotipos y patrones de bandas obtenidos por EGCP (p > 0,05). Conclusiones: Por medio de la detección de marcadores de resistencia y mediante la EGCP, es factible diferenciar la naturaleza de las cepas SAMR de origen clínico. Los resultados obtenidos demuestran la posible transmisión intrahospitalaria de cepas SAMR; así como, su diseminación clonal en los servicios del hospital, particularmente en la UCI, durante el período estudiado.
Assuntos
Humanos , DNA Bacteriano/análise , Staphylococcus aureus Resistente à Meticilina/genética , Antibacterianos/farmacologia , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Eletroforese em Gel de Campo Pulsado , Genótipo , Hospitais Universitários , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Recursos Humanos em Hospital , FenótipoRESUMO
La detección de resistencia a meticilina es complicada debido a la heterogeneidad de su expresión fenotípica; dificultando su detección en el laboratorio, lo que ha conducido al desarrollo de varias técnicas para incrementar su expresión in vitro. A fin de evaluar cuatro técnicas para la detección de resistencia a meticilina: método de difusión del disco con oxacilina (OX, 1 ug) y cefoxitin (FOX, 30 ug); screening test, concentración inhibitoria mínima (CIM) y detección de PBP2a, utilizando la presencia del gen mecA como método de referencia, se procesaron 286 cepas de S. aureus. Se determinó la sensibilidad (SEN), especificidad (ESP), valor predictivo positivo (VPP), valor predictivo negativo (VPN) y eficiencia (EFC) de cada uno de los métodos. Se obtuvo un total de 50 cepas resistentes a oxacilina, PBP2a positivos (mecA positivos). La sensibilidad del disco de OX fue de 99.14 por ciento y la de FOX fue de 100 por ciento. La SEN, VPP, VPN y EFC de los otros métodos fue de 100 por ciento. Todas, a excepción del método de difusión del disco de OX (ESP de 99.14), resultaron 100 por ciento específicos.
Detecting methicillin resistance is complicated due to the heterogeneity of its phenotypic expression, making its detection difficult in the laboratory; this has led to the development of several techniques to increase its expression in vitro. Four techniques for detecting methicillin resistance were evaluated: the disk diffusion method with oxacillin (OX, 1 ug) and cephoxitin (FOX, 30 ug); the screening test, minimum inhibitory concentration (MIC) and detection of PBP2a, using the presence of mecA gen as a reference method; 286 strains of S. aureus, were processed. The sensibility (SEN), specificity (SPE), positive predictive value (PPV), negative predictive value (NPV) and efficiency (EFC) of each method were determined. A total of 50 oxacillin resistant, PBP2a positive (mecA positive) strains were obtained. Sensibility of the OX disk was 99.14 percent; and of the FOX disk was 100 percent. The SEN, PPV, NVP and EFC of the other methods were 100 percent. All the tests, except the OX disk diffusion method (99.14 percent of ESP), were 100 percent specific.