Exploración de características genéticas asociadas al desarrollo del fenotipo de resistencia intermedia heterogénea a vancomicina (hVISA), en aislamientos de Staphylococcus aureus resistente a meticilina (MRSA) causantes de bacteriemia

dc.contributorDíaz Ortiz, Sandra Lorena
dc.contributorPinzón Velasco, Andrés Mauricio
dc.creatorVargas Ardila, Mónica Liliana
dc.date.accessioned2020-07-15T19:26:40Z
dc.date.available2020-07-15T19:26:40Z
dc.date.created2020-07-15T19:26:40Z
dc.date.issued2019
dc.identifierVargas Ardila, Mónica Liliana (2020). Exploración de características genéticas asociadas al desarrollo del fenotipo de resistencia intermedia heterogénea a vancomicina (hVISA), en aislamientos de Staphylococcus aureus resistente a meticilina (MRSA) causantes de bacteriemia. Tesis de Maestría en Ciencias - Microbiología, Universidad Nacional de Colombia - Sede Bogotá.
dc.identifierhttps://repositorio.unal.edu.co/handle/unal/77777
dc.description.abstractVancomycin is the main therapeutic option to treat severe methicillin-resistant Staphylococcus aureus (MRSA) infections; therefore, the emergence of isolates with decreased susceptibility to this antibiotic is a concern. Isolates with heterogeneous intermediate resistance to vancomycin (hVISA) are a clinical and epidemiological challenge, as they are associated with therapeutic and long hospital stays, it is not detectable by standard methodologies, and the genetic mechanism of resistance is not completely understood. However, numerous genes and mutations have been related with this phenotype. For this reason, the aim of this study was to identify genetic characteristics associated with the development of the hVISA phenotype, in MRSA isolates causing bacteremia in 9 Latin American countries. From 538 MRSA isolates, 30 exhibited a positive result for the hVISA phenotype by E-test based methods, however, from these, only 3 were confirmed by population analysis profile/area under the curve (PAP/AUC). Whole genome sequencing was performed to the 30 isolates and amino acid substitutions were identified in 46 proteins associated with intermediate resistance phenotypes (VISA/hVISA) in previous studies. In total, 98 substitutions were found, most related to cell wall biogenesis, DNA/RNA processing, membrane biosynthesis and regulatory systems, from which the most frequent were in VraT (E156G), WalK (L14I), and Atl (Y38H). In addition, changes were also explored in subpopulations of hVISA isolates that grew at concentrations ≥3μg/mL of vancomycin and maintained the hVISA phenotype (no VISA subpopulations). Consistently, no amino acid substitutions were detected other than those found in the isolates from which they were obtained, except for the Y121D change in transport protein PotD. Likewise, a low number of SNPs was detected in the subpopulations when compared with the genomes of the hVISA isolates from which they belonged. In this study, the genetic characterization of hVISA isolates causing bacteremia in patients in South America was performed; previously reported and unreported changes associated with alterations in functions of regulatory systems, cell wall biogenesis, DNA/RNA processing, and membrane biosynthesis were detected, potentially explaining the phenotypic characteristics of hVISA. On another hand, the analysis of hVISA subpopulations showed that the changes detected were consistent with the genomes of the hVISA isolates, suggesting that the mechanisms for the development and regulation of heterogeneous resistance are additional to those reported in the VISA phenotype.
dc.description.abstractVancomicina es la principal opción terapéutica para tratar infecciones severas por Staphylococcus aureus resistente a meticilina (MRSA); por lo tanto, la emergencia de aislamientos con susceptibilidad disminuida a este antibiótico es preocupante. Los aislamientos con resistencia intermedia heterogénea a vancomicina (hVISA) son un reto clínico y epidemiológico, pues están asociados con fallas terapéuticas y largas estancias hospitalarias, no es detectable por metodologías estándar, y el mecanismo genético de resistencia no está completamente elucidado. No obstante, numerosos genes y mutaciones han sido relacionados con este fenotipo. Por esta razón, el objetivo de este estudio fue identificar características genéticas asociadas al fenotipo hVISA, en aislamientos de MRSA causantes de bacteriemia en 9 países latinoamericanos. A partir de 538 aislamientos MRSA, 30 presentaron un resultado positivo para el fenotipo hVISA por métodos basados en E-test, sin embargo, de estos, solo 3 fueron confirmados mediante perfil de análisis poblacional/área bajo la curva (PAP/AUC). A los 30 aislamientos se les realizó secuenciación de genoma completo, con el fin de identificar sustituciones de aminoácidos en 46 proteínas que han sido asociadas a los fenotipos de resistencia intermedia (VISA/hVISA) en estudios previos. En total se encontraron 98 sustituciones, la mayoría relacionadas a biogénesis de pared celular, procesamiento de DNA/RNA, biosíntesis de membrana y sistemas reguladores, de las cuales, las mas comunes se presentaron en VraT (E156G), WalK (L14I) y Atl (Y38H). Adicionalmente, se exploraron cambios en las subpoblaciones de los aislamientos hVISA que crecieron a concentraciones ≥3μg/mL de vancomicina y mantuvieron el fenotipo hVISA (no hubo selección de colonias VISA). De forma consistente, no se detectaron sustituciones de aminoácidos diferentes a las de los aislamientos de donde se obtuvieron, excepto por el cambio Y121D en la proteína de transporte PotD. Así mismo, se detectó un número bajo de SNPs en las subpoblaciones cuando se compararon con los genomas de los aislamientos hVISA de donde provenían. En este estudio se realizó la caracterización genética de aislamientos hVISA causantes de bacteriemia en pacientes de Sur América; se detectaron cambios previamente reportados y cambios no reportados asociados a alteraciones en funciones de sistemas reguladores, biogénesis de pared celular, procesamiento de DNA/RNA, y biosíntesis de membrana, que potencialmente explican las características fenotípicas de hVISA. Por otro lado, el análisis de subpoblaciones de hVISA demostró que los cambios detectados fueron consistentes con los genomas de los aislamientos hVISA, lo cual sugiere que los mecanismos para el desarrollo y regulación de la resistencia heterogénea son adicionales a aquellos reportados en el fenotipo VISA.
dc.languagespa
dc.publisherBogotá - Ciencias - Maestría en Ciencias - Microbiología
dc.publisherUniversidad Nacional de Colombia - Sede Bogotá
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dc.rightsAtribución-NoComercial-SinDerivadas 4.0 Internacional
dc.rightsAcceso abierto
dc.rightshttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rightsinfo:eu-repo/semantics/openAccess
dc.rightsDerechos reservados - Universidad Nacional de Colombia
dc.titleExploración de características genéticas asociadas al desarrollo del fenotipo de resistencia intermedia heterogénea a vancomicina (hVISA), en aislamientos de Staphylococcus aureus resistente a meticilina (MRSA) causantes de bacteriemia
dc.typeOtro


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