• Repositorio Institucional Universidad de Pamplona
  • Tesis de maestría y doctorado
  • Facultad de Ciencias Básicas
  • Maestría en Biología Molecular y Biotecnología
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    dc.contributor.authorChinchilla Cárdenas, Danny.-
    dc.date.accessioned2022-09-24T22:40:17Z-
    dc.date.available2019-11-14-
    dc.date.available2022-09-24T22:40:17Z-
    dc.date.issued2020-
    dc.identifier.citationChinchilla Cárdenas, D. (2019). Implementación de PCR Múltiplex Punto Final y Simplex tiempo real para la Detección de Ehrlichia sp y Hepatozoon canis. [Trabajo de Grado Maestría, Universidad de Pamplona]. Repositorio Hulago Universidad de Pamplona. http://repositoriodspace.unipamplona.edu.co/jspui/handle/20.500.12744/2883es_CO
    dc.identifier.urihttp://repositoriodspace.unipamplona.edu.co/jspui/handle/20.500.12744/2883-
    dc.descriptionEl diagnóstico de las enfermedades infecciosas transmitidas por vectores se realiza de manera rutinaria a través de pruebas serológicas, específicamente pruebas rápidas, incluso en algunos casos se llega a emplear pruebas de Inmunoflorescencia indirecta (IFI) cuantificando títulos de IgG que podrían ser de memoria inmunológica en el caso de Ehrlichia, situación que da posibles falsos positivos (Morales et al, 2015), además del costo elevado. Por otro lado, el diagnóstico de Hepatozoon canis solo se realiza mediante extendidos de sangre periférica, técnica dispendiosa y poco confiable debido a la baja probabilidad de encontrar gametocitos en campos microscópicos en parasitemias bajas (Otranto et al, 2011), es por ello que se desarrolló una técnica de diagnóstico molecular directa altamente sensible y económicamente viable que detecte simultáneamente los dos patógenos más frecuentes en la clínica diaria, las cuales representan gran amenaza contra la vida de las mascotas. Siendo los Hemoparasitos con mayor presencia en Norte de Santander según reporta el Laboratorio MASCOLAB. Por ende se diseñaron por bioinformática, primers específicos que se unen a regiones altamente conservadas de los patógenos en estudio, para luego validar la Reacción en Cadena de la Polimerasa (PCR) según los estándares de Información mínima para experimentos de cuantificación (MIQE) (Bustin et al, 2010), tomando como referencia la tecnología PCR tiempo real cuantitativa, diseñando Sondas de Hibridación. Se tomaron pacientes positivos presuntivamente a Ehrlichia sp, y Hepatozoon canis por extendido de sangre periférica provenientes del área metropolitana de Cúcuta, diagnosticados en el Laboratorio MASCOLAB, a los cuales se les realizó la extracción de ácido dexoxirribonucleico (ADN) por métodos Salting out, y posterior a ello se les realizó la PCR múltiplex punto final y PCR simplex tiempo real sonda taqman para Ehrlichia sp y Hepatozoon canis. El diseño experimental se ensayó con diferentes protocolos de PCR (Top Taq polimerasa Qiagen, Quantinova Syber green Qiagen y Quantinova probe Qiagen, se diseñaron controles internos de amplificación, secuenciación de fragmentos. Se logro amplificar los fragmentos deseados confirmados por secuenciación, obteniendo la PCR múltiplex de detección simultanea de H.canis y Ehrlichia sp. asi mismo, se logro validar el método de extracción de ADN por salting out modificadoes_CO
    dc.description.abstractLa autora no proporciona información sobre este ítem.es_CO
    dc.format.extent100es_CO
    dc.format.mimetypeapplication/pdfes_CO
    dc.language.isoeses_CO
    dc.publisherUniversidad de Pamplona – Facultad de Ciencias Basicas.es_CO
    dc.subjectPCR multiplex punto final.es_CO
    dc.subjectPCR tiempo real sonda taqman.es_CO
    dc.subjectHepatozoon canis.es_CO
    dc.subjectEhrlichia sp.es_CO
    dc.titleImplementación de PCR Múltiplex Punto Final y Simplex tiempo real para la Detección de Ehrlichia sp y Hepatozoon canis.es_CO
    dc.typehttp://purl.org/coar/resource_type/c_bdcces_CO
    dc.date.accepted2019-08-14-
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