Parviz Parvizi
Pasteur Institute of Iran
Title: Evaluating and Setting up a qPCR by High Resolution Melting method for definitely discrimination of Leishmania species by targeting amino acid permease 3 gene
Biography
Biography: Parviz Parvizi
Abstract
Leishmania as protozoan parasites causes major diseases of leishmaniasis in humans in tropical and subtropical regions. In different hosts including humans, clinical samples, rodents and/or other mammals as reservoir hosts and sand flies as vectors, mixed infections, co-infections and different hybrids of Leishmania parasite with different aneuploidy in chromosomes are observed. Differentiate common Old World parasite species and discriminate co-infection with different species. The genetic variation analysis and SNP prediction identify using high resolution melting analysis as a powerful method. For each species, one standard sample was amplified and a recognized region was cloned. Three sets of primer were designed for nuclear gene of amino acid permeases (AAP3) gene and EvaGreen dye mechanism was used and the different temperature of HRM species was optimized. Temperature variation in HRM separated L. major and L. tropica co-infections and their sub-strains. The specific and common primers were separate species and strains by melting temperature analysis. To compare with variety of mitochondrial and nuclear genes, AAP3 gene is more sensitive and specific than other genes for identification of Leishmania parasites. The setup HRM could separate common species of Leishmania parasite and useful in separations intra-stains. Efficiency and regression coefficient reactions for genus and species Leishmania were also validated.