Molecular characterization of Sudanese dermotropic Leishmania parasites

Abstract

Leishmania parasites cause different clinical forms of leishmaniasis constituting major health problems in several countries. Starting from self-healing cutaneous lesions to the fatal, if not treated, visceral form. This study is aimed to characterize and cluster Sudanese dermal Leishmania isolates from different parts of the Sudan. Seventy samples of dermal lesions were collected representing 55 cutaneous ulcers and 15 lesions of PKDL. Of these a total of 57 parasites was successfully cultured, 47 from CL lesions and 10 PKDL lesions. The isolates were characterized based on their kDNA PCR-RFLP patterns and RAPD amplification profiles. PCR amplification of kDNA of Leishmania isolates resulted in amplification of 800 bp in 42 isolates and 700 bp amplicon in 15 isolates, corresponding to L.donovani complex isolates and L.major complex respectively. To investigate the genetic diversity of the isolates, kDNA PCR products were digested with Alu1 restriction enzyme, different RFLP patterns were detected, with noticeable diversity among isolates of the two complexes. Random amplified polymorphic DNA using OP A10 primer resulted amplification of diverse DNA fragments profiles. Phylogenetic trees based on kDNA /RFLP profiles of the two isolates clustered the isolates into two main clusters. Phylogenetic trees based on RAPD profiles of the isolates clustered the isolates into main 5 groups. No correlation between either kDNA /RFLP, or RAPD profiles and the clinical form were detected.