Publication: Estudio y caracterización del microbioma viral en pacientes pediátricos con infección aguda del tracto respiratorio
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2020-04-02
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Universidad Complutense de Madrid
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Abstract
El avance de la tecnología de secuenciación de nueva generación supone una oportunidad para la mejora de la investigación y el diagnóstico, abriendo un horizonte de retos y avances para un laboratorio de referencia. La posibilidad de caracterizar con un único sistema la comunidad viral presente en una muestra biológica y detectar virus, conocidos o no, sin necesidad de cultivar, clonar o restringir la técnica a un virus concreto supone una gran ventaja.El empleo de este sistema en el estudio del tracto respiratorio es un hilo conector que nos lleva desde la puesta a punto de la técnica hasta la comprensión de la ecología de la infección respiratoria y basándose en esto hemos desarrollado nuestros objetivos. El estudio del viroma respiratorio de pacientes pediátricos nos permitió desarrollar los protocolos de enriquecimiento por tres métodos diferentes, de enriquecimiento mediante captura por sondas y la amplificación independiente de secuencia por oligonucleótido único. Estos métodos han sido posteriormente aplicados en el estudio de viroma respiratorio por una parte en niños y en personas adultas, y por otra en hospedadores diferentes al ser humano como son los murciélagos. Este sistema ha sido el método desarrollado y empleado fundamentalmente en este trabajo...
Next-generation sequencing development brings a new horizon of challenges and progress in a reference laboratory providing the opportunity to improve research and diagnostics.The characterization of the viral community with one single method with the ability to find known viruses or not, without the need of cell culturing, cloning or biased molecular methods, is a great advantage. The use of NGS methods is a connecting thread that leads us from the evolvement of the protocols to a better knowledge of respiratory infection. Based on these points the objectives were developed.The pediatric virome study allowed the standardization of the viral enrichment protocol by 3 steps, the virome capture sequencing platform and the sequence independent single primer amplification protocols. These methods were used in the respiratory virome studies of adults and bats.The analysis of the pediatric respiratory virome identified viral pathogens in samples without diagnostic by molecular methods. The absence in the healthy donor samples of the viruses identified in patients with respiratory infection proved the role of these viruses in respiratory infection and characterized the viruses conforming the respiratory virome in health...
Next-generation sequencing development brings a new horizon of challenges and progress in a reference laboratory providing the opportunity to improve research and diagnostics.The characterization of the viral community with one single method with the ability to find known viruses or not, without the need of cell culturing, cloning or biased molecular methods, is a great advantage. The use of NGS methods is a connecting thread that leads us from the evolvement of the protocols to a better knowledge of respiratory infection. Based on these points the objectives were developed.The pediatric virome study allowed the standardization of the viral enrichment protocol by 3 steps, the virome capture sequencing platform and the sequence independent single primer amplification protocols. These methods were used in the respiratory virome studies of adults and bats.The analysis of the pediatric respiratory virome identified viral pathogens in samples without diagnostic by molecular methods. The absence in the healthy donor samples of the viruses identified in patients with respiratory infection proved the role of these viruses in respiratory infection and characterized the viruses conforming the respiratory virome in health...
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Tesis inédita de la Universidad Complutense de Madrid, Facultad de Farmacia, Departamento de Microbiología, leída el 22/11/2019