مقايسه روشهاي مولكولي با ديگر روشهاي متداول آزمايشگاهي در رديابي انگل ليشمانيا در مخازن حيواني بيماري ليشمانيوز جلدي روستايي

Comparision of Molecular Techniques with other Laboratory Methods for Leishmania Detection in ReservoirHosts of Zoonotic CutaneousLeismaniasis

Background and Objective: Zoonotic Cutaneous leismaniasis is one of the important tropical and infectious diseases in Iran and many other countries in the world which sandfies are vectors and rodents blong to gerbilidae family are reservoir hosts. True and indefinite identification of Leishmania parasites in reservoir hosts is important. Specially in reservoir hosts some times there is not any sign of disease and also amount of parasites are few and limited. The objective of this study was to compare molecular techniques with other laboratory methods to detect Leishmania parasites in reservoir hosts of Zoonotic Cutaneous leismaniasis in AghaAli Abbas, Natanz, Isfahan province, Iran. Materials and methods: Two methods tried to extract DNA from ears of rodents and campared with three laborary methods of direct microscopic examination in slides, calutring in NNN and injecting to BALB/c. Using Nested PCR methods amplified a region of the ITS-rDNA gene of Leishmania , which were shown to be species-specific by DNA sequences. Results: From three laborary methods, only one out of 69 rodents was positive by injecting to BALB/c. Direct microscopic examination in slides, calutring in NNN were negative for all 69 rodents. For extarcting DNA, both two methods gave good DNA but ISH-Horovize method was more sensitive. Using Nested PCR of ITS-rDNA gene, gave 10 up to 69 rodents positive. After sequencing, parasites in 9 rodents were Leishmania major and in one rodent was Leishmania tropica. Conclusion: The laboratory methods and molecular techniques with details discused in this paper. Molecular techniques are more sensitive and specific than laboratory methods. Also molecular techniques are more reailable than laboratorry methods to separate and identify different Leishmania species. Because of wild and zoonotic of rodents, it is not easy to sample them by laboratory methods. Sometimes amount of parsites are very low in rodents; these caused negative Leishmania in rodents using laborary methods.