
handle: 10261/229380
[ES] Actualmente, la aparición de resistencia a antibióticos en cepas de Staphylococcus aureus constituye uno de los principales problemas de salud mundial. Por ello, resulta necesario tener un mayor conocimiento de este fenómeno, y buscar estrategias alternativas a los antibióticos para tratar infecciones causadas por este patógeno. En este estudio, se han caracterizado genéticamente, mediante amplificación aleatoria de ADN polimórfico, 67 cepas de S. aureus resistentes a meticilina (MRSA) procedentes del Hospital San Agustín (Avilés, Asturias). Posteriormente, 29 cepas representativas de cada patrón genético y origen clínico fueron sometidas a un ensayo de sensibilidad (reducción de la turbidez) a la endolisina LysRODI (enzibiótico derivado del bacteriófago phiIPLA-RODI). Determinar la actividad específica de dicha endolisina permitió establecer diferencias en la sensibilidad mostrada por las cepas. Seguidamente, se analizó la capacidad de formación de biofilms de las 67 cepas, siendo todas capaces de formarlos, presentando diferencias en cuanto a la biomasa total adherida. Finalmente, se detectaron mediante PCR genes que codifican factores de virulencia y proteínas de superficie en 9 cepas. Los resultados confirmaron la presencia en todas ellas del gen mecA, responsable del fenotipo resistente a meticilina. Además, todas las cepas poseen los genes eno, fnbA, clfA y clfB que codifican adhesinas que reconocen moléculas de la matriz de células del huésped, así como los genes icaA e icaD participantes en la producción de biopelículas. Sin embargo, aunque todas ellas poseen genes que codifican toxinas, existen diferencias en cuáles son los genes presentes. Los resultados obtenidos en este trabajo sugieren que existe una gran variabilidad, genotípica y fenotípica, entre distintas cepas MRSA. Esto supone una mayor complejidad a la hora de seleccionar las estrategias antimicrobianas para su eliminación. Sin embargo, el hecho de que LysRODI mostrara actividad frente a todas las cepas analizadas muestra el potencial de los enzibióticos como una alternativa viable a los antibióticos convencionales.
[EN] Nowadays, the emergence of antibiotic resistance in Staphylococcus aureus strains constitutes one of the main health problems worldwide. For this reason, it is necessary to acquire a greater knowledge of the mechanisms involved in this phenomenon and look for alternative strategies to treat infections by this microorganism. In this study, 67 methicillin-resistant S. aureus (MRSA) strains from San Agustín hospital patients (Avilés, Asturias) were typed by random amplification of polymorphic DNA (RAPD-PCR). Afterwards, 29 strains representing different RAPD patterns and clinical origins were subjected to turbidity assays in order to analyze their susceptibility to the endolysin LysRODI (an enzybiotic derived from phage phiIPLA-RODI). Determination of the specific lytic activity values for the different strains allowed establishing differences in susceptibility amongst them. Besides, all the analyzed strains exhibited varying degrees of biofilm-forming ability. Additionally, the presence of different genes encoding virulence factors, antibiotic resistance determinants and surface proteins was analyzed by PCR in 9 selected strains. The results of these experiments confirmed that all 9 strains harbored the mecA gene, which is responsible for the methicillin-resistance phenotype. Also, all of them carried genes coding for microbial surface components recognizing adhesive matrix molecules, MSCRAMMs (eno, clfA and clfB), as well as genes involved in biofilm formation (icaA and icaD). Finally, all the strains possessed genes involved in toxin production, but not always the same ones. Overall, the results of this work highlight the phenotypic and genotypic diversity of MRSA strains and, consequently, the complexity of developing novel antimicrobial strategies to eliminate this pathogen. Nonetheless, the fact that LysRODI exhibited antimicrobial activity against all the analyzed strains shows the potential of enzybiotics as a viable alternative to conventional antimicrobials.
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