Diseño y evaluación de nuevas técnicas moleculares para la detección y diagnóstico de Leishmania spp. y su aplicación en estudios epidemiológicos

Abstract

The objectives of the present work are: To evaluate the influence of the epidemiological scenario from two endemic areas, Spain and Morocco, on the reliability of the PCR techniques and contribute to the selection of the most efficient one for cutaneous leishmaniosis and typing Leishmania species. To contribute to the improvement in the diagnosis of cutaneous and mucosal leishmaniasis through the establishment of a diagnosis protocol from histological samples. To analyse the composition and distribution of Phlebotomus sergenti mitochondrial lineages in Southwestern Europe using a novel PCR-RFLP for the Cytochrome B population analysis. To investigate Leishmania infantum infection in wild rabbits (Oryctolagus cuniculus) and discuss their potential role as reservoirs in a recognized endemic area with ample epidemiological knowledge, as is the case of Southeastern Spain. The sympatric and syntrophic presence of other kinetoplastid, Trypanosoma nabiasi, was taken into consideration to avoid confusion. To detect and characterize L. infantum infection in wild rodents in an epidemiological study in a well-known focus of canine leishmaniasis due to L. infantum in Southern Spain. To provide an insight into the genetic variability of Trypanosoma nabiasi, its in vitro cultivation and infectivity, and finally investigate the course of infection in its host (O. cuniculus).