Discrimination of leishmania species by arginine permease gene aap3as a suitable molecular tool for detection and genetic proximity analysis

Neda Taleshi,1,* Parviz parvizi,2

Abstract


Introduction

Leishmaniasis presents itself in different clinical manifestations such as: visceral leishmaniasis, cutaneous leishmaniasis, and mucocutaneous leishmaniasis (mcl). in the absent treatment, (vl) is of higher priority than (cl) as it is a fatal diseases. leishmaniasis is part of those disease which still requires improved control tools and should be more focusing on the new methods such as diagnostic tests,new markers. available tools for species identification and phylogenetic analysis include dna sequencing analysis, restriction fragment length polymorphism (rflp) analysis, and pcr techniques. various studies showed that the same target genomic regions can be used to compare distances among species but also to evaluate genetic diversity within species.

Methods

In this study a new set of sensitive and specific primer was designed for aap3 gene and genetic variation was analyzed in clinical samples, serous smears of lesions in humans and rodents suspicion of leishmaniasis and trapped sandflies. sequencing and analysis of their genetic relationship alignment using with software clc workbench 5.5 were performed.

Results

Reported are the results of multidisciplinary studies on leishmania species, which was isolated from the auricular tissues of naturally infected rodents and human and its vectors. designing and testing a new primer set improved sensitivity of the pcr on the mini-exon. in this study a new set of sensitive and specific primer was designed for aap3 gene and genetic variation was analyzed in clinical samples, serous smears of lesions in humans and rodents suspicion of leishmaniasis and trapped sandflies. sequencing and analysis of their genetic relationship alignment using with software clc workbench 5.5 were performed.

Conclusion

Pcr is being used for the diagnosis of parasitic diseases, including leishmaniasis. moreover, parasite of all species of leishmania are morphologically similar, molecular method is required for firmly identification at species level. in term of identification ,priority has been given to marker as feasibility for identification is the highest. various targets have been proposed but in iran for the first time, nuclear gene of amino acid permeases aap3 was tried for firmly identification of leishmania species. the copy number of this gene changes and it have been doubled, can be efficaciously on diffraction of strains. in addition, multicopy genes can be challenging for the importance of this gene in parasite.

Keywords

Aap3,leishmania, pcr