Genetic diversity and phylogenetic analysis of the iranian leishmania parasites based on hsp70 gene pcr-rflp and sequence analysis

Sara Nemati,1,* asghar fazaeli,2 homa hajjaran,3 ali khamesipour,4 mohsen falahati anbaran,5

1. Department of Medical Parasitology and Mycology, School of Medicine, Zanjan University of Medical Sciences, Zanjan 45156
3. Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran
4. Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran 1416613675,
5. School of Biology, University of Tehran, Tehran 14155-6455, Iran

Abstract

Introduction

Despite the broad distribution of leishmaniasis among iranians and animals across the country, little is known about the genetic characteristics of the causative agents

Methods

Applying both hsp70 pcr-rflp and sequence analyses, this study aimed to evaluate the genetic diversity and phylogenetic relationships among leishmania spp. isolated from iranian endemic foci and available reference strains. a total of 36 leishmania isolates from almost all districts across the country were genetically analyzed for the hsp70 gene using both pcr-rflp and sequence analysis. the original hsp70 gene sequences were aligned along with homologous leishmania sequences retrieved from ncbi, and subjected to the phylogenetic analysis. basic parameters of genetic diversity were also estimated

Results

The hsp70 pcr-rflp presented 3 different electrophoretic patterns, with no further intraspecific variation, corresponding to 3 leishmania species available in the country, l. tropica, l. major, and l. infantum. phylogenetic analyses presented 5 major clades, corresponding to 5 species complexes. iranian lineages, including l. major, l. tropica, and l. infantum, were distributed among 3 complexes l. major, l. tropica, and l. donovani. however, within the l. major and l. donovani species complexes, the hsp70 phylogeny was not able to distinguish clearly between the l. major and l. turanica isolates, and between the l. infantum, l. donovani, and l. chagasi isolates, respectively.

Conclusion

our results indicated that both hsp70 pcr-rflp and sequence analyses are medically applicable tools for identification of leishmania species in iranian patients. however, the reduced genetic diversity of the target gene makes it inevitable that its phylogeny only resolves the major groups, namely, the species complexes.

Keywords

Leishmania tropica, leishmania major, leishmania infantum, leishmaniasis, hsp70, phylogenetic analys