Molecular detection of Leishmania DNA in wild-caught sand flies, Phlebotomus and Sergentomyia spp. in northern Iran

Leishmaniasis is currently considered a major health problem in Iran, posing an increasing threat to society's development in various dimensions. This study aimed to detect Leishmania infection in wild-caught sand flies in Sari City, northern Iran. Sand flies were collected using sticky traps, and Leishmania DNA was identified using polymerase chain reaction (PCR) targeting the ITS2-rDNA region, followed by restriction fragment length polymorphism (RFLP) analysis. A total of 138 female sand flies were tested, among which, only 1 specimen of Ph. papatasi (11.11 %) and Ph. major (14.28 %), 4 specimens of Ph. kandelakii (7.27 %) and Se. dentata (8.33 %), and 2 specimens of Se. sintoni (50 %) were naturally infected with L. (L.) major. This was observed in the ITS2 nested-PCR amplification assays where a ∼ 245 bp PCR band was produced. Also, RFLP analysis by Mnl1 revealed the fragments of 55 and 70 and 120 bp for infected sand flies which are characteristic of L. (L.) major. Most of the sand flies were unfed, collected during warm season, found indoor. This study reperesents the first molecular detection of L. (L.) major in wild-caught sand flies, specifically in Ph. papatasi in this region, as well as Ph. kandelakii and Ph. major in Iran and even the world.