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. 2016 Jan 5;10(2):141-7.
eCollection 2016 Jun.

Molecular Detection of Leishmania major and L. turanica in Phlebotomus papatasi and First Natural Infection of P. salehi to L. major in North-East of Iran

Affiliations

Molecular Detection of Leishmania major and L. turanica in Phlebotomus papatasi and First Natural Infection of P. salehi to L. major in North-East of Iran

Sayena Rafizadeh et al. J Arthropod Borne Dis. .

Abstract

Background: Leishmaniasis is an important public health disease in many developing countries as well in Iran. The main objective of this study was to investigate on leishmania infection of wild caught sand flies in an endemic focus of disease in Esfarayen district, north east of Iran.

Methods: Sand flies were collected by sticky papers and mounted in a drop of Puri's medium for species identification. Polymerase chain reaction techniques of kDNA, ITS1-rDNA, followed by restriction fragment length polymorphism were used for identification of DNA of Leishmania parasites within infected sand flies.

Results: Among the collected female sand flies, two species of Phlebotomus papatasi and Phlebotomus salehi were found naturally infected with Leishmania major. Furthermore, mixed infection of Leishmania turanica and L. major was observed in one specimen of P. papatasi. Sequence analysis revealed two parasite ITS1 haplotypes including three L. major with accession numbers: KJ425408, KJ425407, KM056403 and one L. turanica. (KJ425406). The haplotype of L. major was identical (100%) to several L. major sequences deposited in GenBank, including isolates from Iran, (Gen Bank accession nos.AY573187, KC505421, KJ194178) and Uzbekistan (Accession no.FN677357).

Conclusion: To our knowledge, this is the first detection of L. major within wild caught P. salehi in northeast of Iran.

Keywords: Iran; L. turanica; Leishmania major; P. salehi; Phlebotomus papatasi.

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Figures

Fig. 1.
Fig. 1.
kDNA nested PCR amplification (560 bp). L. major in P. papatasi (Lane B, C), L. major in P. salehi (LaneD), Mixed infection of L. major and L. turanica in P. papatasi (lane A), Positive control of L. tropica (Lane P3, 720 bp), Positive control of L. major (Lane P2), Positive control of L. infantum (680 bp, LaneP1), Negative control (Lane N) and (L) 100 bp molecular weight marker (Fermentase)
Fig. 2.
Fig. 2.
ITS1 amplification of L. major in P. papatasi (Lane B,C) and P. salehi (Lane D), Mixed infection of L. major and L. turanica in P. papatasi (lane A), positive control of L. major (P), Negative control (Lane N) and (L)100 bp molecular weight marker (Fermentase)
Fig. 3.
Fig. 3.
PCR-RFLP analysis of ITS1 region for identification of Leishmania species using HaeIII. (L) 100 bp molecular weight marker, (P), positive control of L. major, (N) negative control, (A,B,C) samples of infected P. papatasi to L. major, (D) Infected P. salehi to L. major

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