Molecular typing of Leishmania major parasites by targeting Microsatellite gene of main reservoir hosts in Khuzestan province

Summary and Objective: Leishmania major is a significant causative agent of zoonotic cutaneous leishmaniasis (ZCL) in Khuzestan province and Tatera indica species has been incriminated as main reservoir hosts in this region. Only molecular methods are able to identify Leishmania parasites firmly in rodents. Whereas ordinary laboratory methods have not yet been enough to detect of Leishmania, therefore, theLeishmania major parasites have been typed by targeting Microsatellite gene. Investigation method: Tatera indica rodents were captured using wooden and live wiry traps then were identified bydiagnostic keys and molecular tools. Smears of each ear were prepared from serous ofscratching ears. DNAs were extracted by Genet Bio kit. In addition, Microsatellite gene was amplified by Nested PCR and measuring band size. Thereby, Leishmania species has been surely approved by their sequences. Results: In this investigation, Tatera indica rodents were discriminated from other gerbils. L. major was diagnosed decisively after DNA extracting and amplifying of relevant gene in this rodent. Using nucleotide sequencing of the L. major gene were performed by SequencherTm 4.1.1 software, the sequences were edited, aligned and compared with the registered microsatellite gene of L. major in GenBank in a case of similarity and homology. All sequences (Forward & Reverse) were made to a single network (contig) and analysed systematically with MEGA and PAUP software which were employed for phylogenetic analysis. The isolated samples of Leishmania major from this rodent (Tatera indica) were contained one common and/or general haplotype and one new unique haplotype which are being reported for the first time.

Keywords:

Leishmania major , rural leishmaniasis , Microsatellite gene , Molecular typing , Khuzestanprovince