Detección del Genoma del Papilomavirus humano aplicando la técnica Reacción en cadena de la Polimerasa en tiempo real.
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2008
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El Papilomavirus humano (HPV por sus siglas en inglés) pertenece al Género Papilomavirus de la familia Papoviridae y son virus sin envoltura. En particular la familia Papoviridae incluye virus que provocan evidentes afecciones clínicas no solo en humanos sino también en animales. Desde la década de los 90 varios estudios con el soporte de la tecnología molecular han demostrado que la infección con genotipos del Papilomavirus humano de alto riesgo es una causa bien establecida del desarrollo de cáncer cérvico uterino [1, 3, 30], patología que constituye el 51% de todos los cánceres detectados en la población femenina que acude al ION SOLCA-Guayaquil. Técnicas de detección temprana molecular del HPV como la Reacción en Cadena de la Polimerasa convencional (Polimerase Chain Reaction, PCR), PCR en tiempo real (real-time PCR) entre otras, se están convirtiendo en importantes herramientas de los protocolos de prevención y manejo del cáncer cérvico uterino in situ. El presente trabajo de tesis busca detectar el genoma del Papilomavirus humano mediante la técnica de la PCR en tiempo real en pacientes sospechosas o ya diagnosticadas con infección por HPV y comparar su especificidad y sensibilidad con la técnica diagnóstica de histopatología. Se analizó un total de 99 muestras obtenidas de 52 pacientes sospechosas o con diagnóstico de infección por HPV, divididas en dos grupos de análisis para comparar técnicas de extracción de ADN. La extracción mediante el uso del Kit Wizard de Promega dio mejores resultados en la amplificación por PCR en tiempo real, al obtener ADN de mayor pureza comparado con el método Invitrogen –Innogenetics. Al evaluar la calidad de la detección por PCR en tiempo real nos dio cómo resultado un 95.23% de sensibilidad y un 11.11% de especificidad
The human papillomavirus (HPV acronym) belongs to the family Papillomavirus Genre Papoviridae and are non-enveloped viruses. In particular Papoviridae family includes viruses that cause obvious clinical conditions not only in humans but also in animals. From the 90's several studies supported by molecular technology have shown that infection with human papillomavirus genotypes is a high risk of developing well-established cause of cervical cancer [1, 3, 30], pathology is 51% of all cancers detected in women who go to ION SOLCA-Guayaquil. Molecular techniques for early detection of HPV as Chain Reaction conventional Polymerase (Polymerase Chain Reaction, PCR), real-time PCR (real-time PCR) among others, are becoming important tools for the prevention and management protocols cervical cancer in situ. This thesis seeks to detect the human papillomavirus genome using the technique of real-time PCR in patients suspected or already diagnosed with HPV infection and its specificity and sensitivity compared with histopathology diagnostic technique. We analyzed a total of 99 samples obtained from 52 patients suspected or diagnosed with HPV infection, divided into two groups of analysis to compare DNA extraction techniques. Extraction Kit using the Promega Wizard gave better results in the PCR amplification in real time, to obtain DNA of higher purity compared to the method-Innogenetics Invitrogen. To evaluate the quality of the detection by real-time PCR gave 95.23% resulting in a sensitivity and specificity
The human papillomavirus (HPV acronym) belongs to the family Papillomavirus Genre Papoviridae and are non-enveloped viruses. In particular Papoviridae family includes viruses that cause obvious clinical conditions not only in humans but also in animals. From the 90's several studies supported by molecular technology have shown that infection with human papillomavirus genotypes is a high risk of developing well-established cause of cervical cancer [1, 3, 30], pathology is 51% of all cancers detected in women who go to ION SOLCA-Guayaquil. Molecular techniques for early detection of HPV as Chain Reaction conventional Polymerase (Polymerase Chain Reaction, PCR), real-time PCR (real-time PCR) among others, are becoming important tools for the prevention and management protocols cervical cancer in situ. This thesis seeks to detect the human papillomavirus genome using the technique of real-time PCR in patients suspected or already diagnosed with HPV infection and its specificity and sensitivity compared with histopathology diagnostic technique. We analyzed a total of 99 samples obtained from 52 patients suspected or diagnosed with HPV infection, divided into two groups of analysis to compare DNA extraction techniques. Extraction Kit using the Promega Wizard gave better results in the PCR amplification in real time, to obtain DNA of higher purity compared to the method-Innogenetics Invitrogen. To evaluate the quality of the detection by real-time PCR gave 95.23% resulting in a sensitivity and specificity
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DETECCIÓN DEL GENOMA, PAPILOMAVIRUS HUMANO, TÉCNICA REACCIÓN EN CADENA