Thomas, Megan and Bates, Paul (2018) Identification of Leishmania tropica and Leishmania aethiopica by DNA sequencing and the detection of Leishmania RNA Virus. Masters thesis, Lancaster University.
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Abstract
Leishmaniasis is a neglected tropical disease and a major cause of morbidity and mortality in impoverished countries. Leishmaniasis is a global disease and is clinically and epidemiologically diverse, with many different species of Leishmania affecting humans. The aim of this study was to investigate the classification of L. tropica and L. aethiopica as different species, and to check for the presence of LRV in their isolates. L. tropica and L. aethiopica are closely related species of Leishmania that infect humans; and there is some debate over whether they are indeed different species. The Leishmania RNA Virus is a double stranded RNA virus which infects Leishmania and is thought to increase the pathology of the disease and exacerbating mucosal developments. The methods used in this study were polymerase chain reaction and sequencing for the ITS1 gene and a dot blot to detect the presence of LRV. The results of the PCR were used for identification, the formation of phylogenetic trees, and a distance pairwise matrix. The PCR results supported the classification of L. tropica and L. aethiopica as different species, supporting previous results also used for the analysis. The results of the dot blot found LRV presence in the L. aethiopica isolates but no evidence of the virus in L. tropica. The identification of an infecting species of Leishmania and the clinical outcomes it can cause will help provide appropriate and affective treatment. The findings of this study also demonstrate the importance of species identification and the methods used.