{"id":60092,"date":"2018-03-09T22:47:56","date_gmt":"2018-03-09T22:47:56","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/desarrollo-y-estandarizacion-de-tecnicas-de-pcr-multiple-para-la-deteccion-de-patogenos-bacterianos-de-importancia-en-acuicultura\/"},"modified":"2018-03-09T22:47:56","modified_gmt":"2018-03-09T22:47:56","slug":"desarrollo-y-estandarizacion-de-tecnicas-de-pcr-multiple-para-la-deteccion-de-patogenos-bacterianos-de-importancia-en-acuicultura","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/microbiologia\/desarrollo-y-estandarizacion-de-tecnicas-de-pcr-multiple-para-la-deteccion-de-patogenos-bacterianos-de-importancia-en-acuicultura\/","title":{"rendered":"Desarrollo y estandarizaci\u00f3n de t\u00e9cnicas de pcr m\u00faltiple para la detecci\u00f3n de pat\u00f3genos bacterianos de importancia en acuicultura"},"content":{"rendered":"<h2>Tesis doctoral de <strong>  Mata Mart\u00edn Ana Isabel <\/strong><\/h2>\n<p>El estudio se ha basado en el dise\u00f1o de diferentes ensayos de pcr m\u00faltiple, como t\u00e9cnica de diagn\u00f3stico alternativa a los m\u00e9todos microbiol\u00f3gicos tradicionales, para la detecci\u00f3n simult\u00e1nea de varios pat\u00f3genos bacterianos de peces.  en una primera parte, se ha desarrollado un ensayo de pcr m\u00faltiple para los principales pat\u00f3genos bacterianos causantes de las estreptococosis de agua templada en peces (lactococcus garvieae, streptococcis iniae, streptococcus parauberis y streptococcus difficilis), enfermedades que causan cada vez mayores p\u00e9rdidas econ\u00f3micas en la acuicultura espa\u00f1ola. En la puesta a punto de este ensayo de pcr se comprob\u00f3, en primer lugar, la especificidad de los oligonucle\u00f3tidos seleccionados, descritos en la bibliograf\u00eda, a partir de lo cual se procedi\u00f3 al dise\u00f1o de nuevos indicadores para s. Iniae, tomando como diana molecular el gen icto de este pat\u00f3geno, que codifica para la enzima lactato oxidasa. Los oligonucle\u00f3tidos dise\u00f1ados (denominados lox-1 y lox-2) amplifican de forma espec\u00edfica un fragmento de 870 pb del gen icto de s. Iniae. El l\u00edmite de detecci\u00f3n de esta pcr individual para la identificaci\u00f3n de s. Iniae, partiendo de adn purificado, fue de 25 pg de adn, y de 62-31 c\u00e9lulas por mezcla de reacci\u00f3n, partiendo de cultivos puros bacterianos. La pcr se ensay\u00f3 tambi\u00e9n sobre tejidos de peces inoculados artificialmente, obteni\u00e9ndose, en este caso, un l\u00edmite de detecci\u00f3n de 4 x 10elevado a 3 c\u00e9lulas\/g de ri\u00f1\u00f3n y de 5 x 10elevado a 3 c\u00e9lulas\/g de h\u00edgado o cerebro.  las cuatro parejas que finalmente se emplearon en la pcr m\u00faltiple desarrolla para la detecci\u00f3n simult\u00e1nea de los principales pat\u00f3genos productores de estreptococosis, fueron: plg-1\/plg-2 para l. Garvieae, lox-1\/lox-2, dise\u00f1ados en este estudio, para s. Iniae, spa-2152\/spa-2870 para s. Parauberis y sdi-61\/sdi-252 para s. Difficillis. La combinaci\u00f3n de estas cuatro parejas de oligonucle\u00f3tidos en un mismotubo de reacci\u00f3n dio lugar a la ampli<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Desarrollo y estandarizaci\u00f3n de t\u00e9cnicas de pcr m\u00faltiple para la detecci\u00f3n de pat\u00f3genos bacterianos de importancia en acuicultura<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Desarrollo y estandarizaci\u00f3n de t\u00e9cnicas de pcr m\u00faltiple para la detecci\u00f3n de pat\u00f3genos bacterianos de importancia en acuicultura <\/li>\n<li><strong>Autor:<\/strong>\u00a0  Mata Mart\u00edn Ana Isabel <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Complutense de Madrid<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 19\/07\/2007<\/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>garayz\u00e1bal Fern\u00e1ndez Jos\u00e9 Francisco Fern\u00e1ndez<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: lucas Dom\u00ednguez rodr\u00edguez <\/li>\n<li>Mar\u00eda isabel Gonz\u00e1lez alonso (vocal)<\/li>\n<li> Latre cequiel Mar\u00eda victoria (vocal)<\/li>\n<li>ignacio De blas giral (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Mata Mart\u00edn Ana Isabel El estudio se ha basado en el dise\u00f1o de diferentes ensayos de pcr [&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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