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Circulación Del Virus Influenza B En Uruguay: Epidemiología Y Diversidad Genética

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Circulación Del Virus Influenza B En Uruguay: Epidemiología Y Diversidad Genética UNIVERSIDAD DE LA REPÚBLICA PEDECIBA BIOLOGÍA – Opción MICROBIOLOGÍA TESIS DE MAESTRÍA CIRCULACIÓN DEL VIRUS INFLUENZA B EN URUGUAY: EPIDEMIOLOGÍA Y DIVERSIDAD GENÉTICA María José Rivas Amiassorho Marzo 2020 Tutor: Dra. Natalia Goñi Co-Tutor: Dra. Dora Ruchansky Lugar de trabajo: Centro Nacional de Referencia de Influenza. Laboratorio de Virología del Departamento de Laboratorios del Ministerio de Salud. 1 Índice Agradecimientos ........................................................................................................................................................................ 4 Resumen ..................................................................................................................................................................................... 5 Listado de abreviaturas ............................................................................................................................................................. 6 1. Clasificación y nomenclatura ........................................................................................................................................... 7 2. Morfología y estructura del virus ..................................................................................................................................... 7 3. Ciclo viral ............................................................................................................................................................................ 9 3.1. Adsorción, entrada y transporte de los RNPv ............................................................................................. 9 3.2. Transcripción y síntesis proteínas virales .................................................................................................. 12 3.3. Replicación ..................................................................................................................................................... 13 3.4. Empaquetamiento y ensamble .................................................................................................................... 13 3.5. Brotamiento y liberación ............................................................................................................................... 14 4. Genoma viral.................................................................................................................................................................... 14 5. Glicosilación de los virus Influenza............................................................................................................................... 17 6. Hemaglutinina .................................................................................................................................................................. 20 6.1. Sitios antigénicos........................................................................................................................................... 21 6.2. Sitio de unión al receptor .............................................................................................................................. 24 7. Neuraminidasa ................................................................................................................................................................ 24 8. Ecología ............................................................................................................................................................................ 26 9. Manifestaciones clínicas de la infección causada por el VIB y epidemiología ...................................................... 26 10. Estacionalidad ................................................................................................................................................................. 29 11. Evolución .......................................................................................................................................................................... 30 11.1. Linajes del virus Influenza B ........................................................................................................................ 31 11.2. Patogenicidad ................................................................................................................................................ 33 11.3. Balance HA:NA .............................................................................................................................................. 34 11.4. Evolución de glicoproteínas de superficie ................................................................................................. 35 11.5. Reordenamientos .......................................................................................................................................... 38 12. Inmunidad......................................................................................................................................................................... 39 13. Tratamiento y prevención .............................................................................................................................................. 42 13.1. Vacunas .......................................................................................................................................................... 42 13.2. Nuevas tecnologías de suministro y fabricación ....................................................................................... 47 13.3. Tratamiento y profilaxis con drogas antivirales ......................................................................................... 48 14. Resistencia entre los VIB a los INA .............................................................................................................................. 49 15. Vigilancia .......................................................................................................................................................................... 51 16. Diagnóstico ...................................................................................................................................................................... 52 17. Objetivos........................................................................................................................................................................... 54 17.1. Objetivo general............................................................................................................................................. 54 17.2. Objetivos específicos .................................................................................................................................... 54 2 18. Materiales y métodos ..................................................................................................................................................... 55 18.1. Vigilancia Influenza y recolección de muestras ........................................................................................ 55 18.2. Métodos estadísticos .................................................................................................................................... 55 18.3. Detección molecular de VIB......................................................................................................................... 55 18.3.1. Extracción de ARN y ensayos RT-PCR en tiempo real ........................................................................... 55 18.3.2. PCR Convencional y electroforesis ............................................................................................................ 56 18.3.3. Purificación de bandas y secuenciación .................................................................................................... 57 18.3.4. Datos de secuencias y análisis filogenéticos de los genes HA y NA de VIB ....................................... 57 18.3.5. Variaciones en secuencias aminoacídicas y sitios de glicolisalición ..................................................... 58 19. Resultados ....................................................................................................................................................................... 59 19.1. Detección molecular de los VIB circulantes en Uruguay ......................................................................... 59 19.1.1. Análisis Filogenético. Gen HA ..................................................................................................................... 59 19.1.2. Análisis filogenéticos. Gen NA .................................................................................................................... 63 19.2. Sustitución de aminoácidos en la región HA1 ........................................................................................... 66 19.2.1. Sitio unión al receptor y sitios de glicosilación .......................................................................................... 68 19.3. Sustitución de aminoácidos en la NA ......................................................................................................... 69 19.3.1. Sustituciones en sitio activo, sitios de glicosilación y sitios de resistencia ........................................... 72 19.4. Distribución de edades según tipo de vigilancia ....................................................................................... 72 19.5. Prevalencia de los virus influenza B en Uruguay ..................................................................................... 73 19.6. Interacción
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