Caracterización molecular y evaluación de la susceptibilidad de especies de Leishmania, circulantes en Bolivia, a nuevos fármacos y formulaciones
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2015
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23/06/2014
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Universidad Complutense de Madrid
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La leishmaniosis es una enfermedad causado por más de 20 especies del género Leishmania que causan diferentes patologías conocidas como leishmaniosis visceral (LV), leishmaniosis cutáneo (LC) y leishmaniosis mucocutáneo (LMC). Es una enfermedad prevalente en 98 países y afecta a más de 1.3 millones de personas. En Latinoamérica, es una enfermedad endémica y Bolivia registró la mayor tasa de incidencia en 2006 (33 casos por cada 100.000 habitantes), de los cuales más del 50% se localiza en la región subandina del Departamento de La Paz. La gravedad de la enfermedad se asocia a la variabilidad intrínseca de Leishmania, los fracasos terapéuticos, la resistencia adquirida y los efectos adversos de los tratamientos convencionales. En este contexto, los expertos de la Organización Mundial de la Salud, han exhortado a las instituciones públicas y privadas a aunar esfuerzos en la identificación de las especies circulantes en cada región, priorizar las investigaciones en la búsqueda de alternativas terapéuticas e investigar los factores asociados a las fallos terapéuticos y de resistencia. La presente memoria de tesis doctoral, fue diseñada para identificar las especies de Leishmania circulantes en el Departamento de La Paz (Bolivia) y evaluar su susceptibilidad farmacológica in vitro. Además, se incorpora un tercer objetivo dirigido a la búsqueda de alternativas terapéuticas, claves para el futuro tratamiento de la leishmaniosis...
Leishmaniosis is an infectious disease caused by over 20 Leishmania species resulting in multiple clinical forms of the disease: visceral (VL), cutaneous (CL) and mucocutaneous (MCL). Leishmaniosis are prevalent in 98 countries and affect more than 1.3 million of people worldwide. Is an endemic disease in Latin America and Bolivia has the highest incidence of CL, with 33 cases per 100,000 population reported in 2006. More than 50% of these cases are localized in subandean region of the department La Paz being one of the biggest affected areas in the country. Conventional drug therapeutic failure and appearance of resistance can be associated to the high variability of Leishmania spp could be lead in severity of the disease. The WHO (World Health Organization) is actively promoting the joining of public and private institutions in Bolivia with the aim of discovering new cost-‐ effective treatments and identifying which factors are related to drug failure and resistance. The aim of this Thesis was to identify all the different Leishmania spp. localised in the Department of La Paz (Bolivia) and assess their in vitro drug susceptibility. Also, the development of new cost-‐effective treatments was studied. 55 Leishmania isolates from patients who had been diagnosed with cutaneous and mucocutaneous Leishmaniosis by the Parasitology laboratory of National Institute of Laboratory Health (INLASA) in Bolivia were obtained. Parasites were kept in culture by periodically passaging. DNA was purified and analysed using three molecular markers: internal transcribed spacer (ITS-‐1), heat shock protein (HSP70-‐ I) and cytochrome b gene (cyt-‐b). DNA was amplified using PCR and the obtained fragments were digested by restriction endonucleases. PCR products of cyt-‐b also were sequenced...
Leishmaniosis is an infectious disease caused by over 20 Leishmania species resulting in multiple clinical forms of the disease: visceral (VL), cutaneous (CL) and mucocutaneous (MCL). Leishmaniosis are prevalent in 98 countries and affect more than 1.3 million of people worldwide. Is an endemic disease in Latin America and Bolivia has the highest incidence of CL, with 33 cases per 100,000 population reported in 2006. More than 50% of these cases are localized in subandean region of the department La Paz being one of the biggest affected areas in the country. Conventional drug therapeutic failure and appearance of resistance can be associated to the high variability of Leishmania spp could be lead in severity of the disease. The WHO (World Health Organization) is actively promoting the joining of public and private institutions in Bolivia with the aim of discovering new cost-‐ effective treatments and identifying which factors are related to drug failure and resistance. The aim of this Thesis was to identify all the different Leishmania spp. localised in the Department of La Paz (Bolivia) and assess their in vitro drug susceptibility. Also, the development of new cost-‐effective treatments was studied. 55 Leishmania isolates from patients who had been diagnosed with cutaneous and mucocutaneous Leishmaniosis by the Parasitology laboratory of National Institute of Laboratory Health (INLASA) in Bolivia were obtained. Parasites were kept in culture by periodically passaging. DNA was purified and analysed using three molecular markers: internal transcribed spacer (ITS-‐1), heat shock protein (HSP70-‐ I) and cytochrome b gene (cyt-‐b). DNA was amplified using PCR and the obtained fragments were digested by restriction endonucleases. PCR products of cyt-‐b also were sequenced...
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Tesis inédita de la Universidad Complutense de Madrid, Facultad de Farmacia, Departamento de Parasitología, leída el 23/06/2014