{"id":96191,"date":"2018-03-11T10:16:21","date_gmt":"2018-03-11T10:16:21","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/sensores-electroqua%c2%admicos-de-adn-para-la-deteccion-de-legionella-pneumophila\/"},"modified":"2018-03-11T10:16:21","modified_gmt":"2018-03-11T10:16:21","slug":"sensores-electroqua%c2%admicos-de-adn-para-la-deteccion-de-legionella-pneumophila","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/quimica-analitica\/sensores-electroqua%c2%admicos-de-adn-para-la-deteccion-de-legionella-pneumophila\/","title":{"rendered":"Sensores electroqu\u00edmicos de adn para la detecci\u00f3n de legionella pneumophila"},"content":{"rendered":"<h2>Tesis doctoral de <strong> Rebeca Miranda Castro <\/strong><\/h2>\n<p>En este trabajo, se ha desarrollado un genosensor electroqu\u00edmico de hibridaci\u00f3n con formato s\u00e1ndwich y amplificaci\u00f3n enzim\u00e1tica para secuencias especificas de legionella pneumophila, en el que se emplea como sonda de captura una secuencia de adn con forma de horquilla. Se ha demostrado que el empleo de este tipo de secuencias sonda da lugar a mejoras de sensibilidad y, especialmente, de selectividad en comparaci\u00f3n con las cl\u00e1sicas sondas lineales. El genosensor que mejores prestaciones anal\u00edticas ofrece se obtiene por quimisorci\u00f3n sobre electrodos de oro de monocapas binarias oligonucle\u00f3tido estructurado tiolado en el extremo 5\u00c2\u00bfmercaptohexanol. En el ensayo s\u00e1ndwich se emplea como secuencia de \u00e1cido nucleico informadora un oligonucle\u00f3tido modificado con el enzima fosfatasa alcalina a trav\u00e9s del enlace biotina-estreptavidina. Con este esquema de dise\u00f1o y un formato de detecci\u00f3n voltamperom\u00e9trica se obtienen excelentes caracter\u00edsticas de detectabilidad, con un l\u00edmite de detecci\u00f3n de 340 pm y un rango lineal hasta 2 \u00c2\u00b5m. El sensor presenta adem\u00e1s extraordinaria selectividad, ya que permite discriminar entre secuencias especificas de l. Pneumophila y l. Longbeachae o l. Micdadei sin necesidad de establecer condiciones dr\u00e1sticas de trabajo. El ensayo optimizado ha sido adaptado a un dispositivo desechable utilizando electrodos serigrafiados. Asimismo, este esquema de detecci\u00f3n ha sido integrado en un sistema microflu\u00eddico con detecci\u00f3n electroqu\u00edmica, reduciendo considerablemente el tiempo de an\u00e1lisis y el consumo de reactivos. El genosensor electroqu\u00edmico se ha aplicado a la detecci\u00f3n y cuantificaci\u00f3n de legionella pneumophila en muestras de agua contaminada, utilizando una etapa previa de amplificaci\u00f3n mediante pcr. Este ensayo permite detectar l. Pneumophila de manera simple y r\u00e1pida en muestras que contienen solamente 10e2 copias del genoma de la bacteria y distinguir entre \u00e9stas y 10e3 o 10e4 copias del pat\u00f3geno, valores l\u00edmite que se recogen en la legislaci\u00f3n ambiental vigente para determinar el tipo de acciones correctivas en instalaciones con riesgo de proliferaci\u00f3n de l. Pneumophila.<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Sensores electroqu\u00edmicos de adn para la detecci\u00f3n de legionella pneumophila<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Sensores electroqu\u00edmicos de adn para la detecci\u00f3n de legionella pneumophila <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Rebeca Miranda Castro <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Oviedo<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 25\/09\/2009<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h3>Direcci\u00f3n y tribunal<\/h3>\n<ul>\n<li><strong>Director de la tesis<\/strong>\n<ul>\n<li>Arturo Jos\u00e9 Miranda Ordieres<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: paulino Tu\u00f1\u00f3n blanco <\/li>\n<li>beatriz L\u00f3pez ruiz (vocal)<\/li>\n<li>giovanna Marrazza (vocal)<\/li>\n<li>Mar\u00eda   encarnacion Lorenzo abad (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Rebeca Miranda Castro En este trabajo, se ha desarrollado un genosensor electroqu\u00edmico de hibridaci\u00f3n con formato s\u00e1ndwich [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center 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