Molecular detection and conventional identification of leishmania species in reservoir hosts of zoonotic cutaneous leishmaniasis in fars province, south of Iran

ABSTRACT

The objectives of our research were to search for Leishmania species in rodents in Fars province, south of Iran, and to compare molecular with conventional methods for detecting these parasites. Rodents were captured using live traps and screened for Leishmania species using molecular and conventional methods, including the taking of smears from each ear. Nested PCR was employed to detect Leishmania in rodents by amplifying a region of the ribosomal RNA amplicon of Leishmania [ITS1- 5.8S rRNA-ITS2] that is species-specific by DNA sequence. Totally, 122 rodents were captured. Leishmania parasites were detected using the nested PCR and three conventional methods [direct smear, NNN culture and Balb/C inoculation. 41 [33.6%] out of 122 rodents had Leishmania infections [34 Meriones lybicus and 7 M. persicus]. All PCR products of the ITS-rDNA gene were sequenced. Sequence analysis revealed that 28 out of 41 positive samples were Leishmania major. Thirteen sequences were unreadable and therefore not identified. At least two gerbil species common in Fars ZCL foci, M. lybicus and M. persicus, are acquiring infections of L. major and may be reservoir hosts of one predominant parasite haplotype. Most infections were detected molecularly not by conventional methods, because most rodents died in the traps