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Objective:To identify the Leishmaniaspecies in infected sand flies byReal-timePCR coupled withHRM analysis.Methods:Real-timePCR coupled withHRM analysis targeting the first intal transcribed spacer(ITS1) of nuclear ribosomalDNA as the genetic marker was used to identify and distinguish Leishmania species in sand flies specimens.Results:Three out of115 females ofPhlebotomus sergenti(P. sergenti)(2.6%) were positive toLeishmania tropica(L. tropica). Conclusions:This is the first report onP. sergenti as the main and proven vector of anthroponitic cutaneous leishmaniasis inDehbakriCounty usingReal-timePCR coupled withHRM analysis. This method is rapid, sensitive and specific for diagnosing of parasites in infectedSand flies and ideal for large scale genotyping projects.