title: Identification of Tunisian Leishmania spp. by PCR amplification of cysteine proteinase B (cpb) genes and phylogenetic analysis.
abstract: Discrimination of the Old World Leishmania parasites is important for diagnosis and epidemiological studies of leishmaniasis. We have developed PCR assays that allow the discrimination between Leishmania major, Leishmania tropica and Leishmania infantum Tunisian species. The identification was performed by a simple PCR targeting cysteine protease B (cpb) gene copies. These PCR can be a routine molecular biology tools for discrimination of Leishmania spp. from different geographical origins and different clinical forms. Our assays can be an informative source for cpb gene studying concerning drug, diagnostics and vaccine research. The PCR products of the cpb gene and the N-acetylglucosamine-1-phosphate transferase (nagt) Leishmania gene were sequenced and aligned. Phylogenetic trees of Leishmania based cpb and nagt sequences are close in topology and present the classic distribution of Leishmania in the Old World. The phylogenetic analysis has enabled the characterization and identification of different strains, using both multicopy (cpb) and single copy (nagt) genes. Indeed, the cpb phylogenetic analysis allowed us to identify the Tunisian Leishmania killicki species, and a group which gathers the least evolved isolates of the Leishmania donovani complex, that was originated from East Africa. This clustering confirms the African origin for the visceralizing species of the L. donovani complex.
authors.affiliation: Laboratoire d'Epidémiologie et d'Ecologie Parasitaire, Institut Pasteur de Tunis, Tunis-Belvédère, Tunisia.
authors.forename: Melek
authors.initials: M
authors.lastname: Chaouch
authors.forename: Akila
authors.initials: A
authors.lastname: Fathallah-Mili
authors.forename: Mehdi
authors.initials: M
authors.lastname: Driss
authors.forename: Ramzi
authors.initials: R
authors.lastname: Lahmadi
authors.forename: Chiraz
authors.initials: C
authors.lastname: Ayari
authors.forename: Ikram
authors.initials: I
authors.lastname: Guizani
authors.forename: Moncef
authors.initials: M
authors.lastname: Ben Said
authors.forename: Souha
authors.initials: S
authors.lastname: Benabderrazak
mesh_terms: D016000:Cluster Analysis; D057056:Cysteine Proteases / Q000235:genetics*; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D007891:Leishmania / Q000145:classification* / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D007896:Leishmaniasis / Q000469:parasitology; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics; D014416:Tunisia
chemical_list: D016054:DNA, Protozoan; D017855:Transferases (Other Substituted Phosphate Groups); C023222:UDPacetylglucosamine-dolichyl-phosphate acetylglucosamine-1-phosphate transferase; D057056:Cysteine Proteases