Molecular detection of Leishmania species in Sand Flies by PCR-RFLP technique in refugee camps

[Detecção molecular de espécies de Leishmania em flebotomíneos por meio da técnica PCR-RFLP em campos de refugiados]

ABSTRACT

Leishmaniasis is one of the most important health dilemmas facing the World Health Organization (WHO), due to it being widespread and the great diversity of sand flies that transmit it. This study aimed to detect the presence of Leishmania parasites in the sand flies spread in Refugee camps by PCR- RLFP technique. A total of 437 sandflies were collected and classified into two species Phlebotomus papatasi and Phlebotomus sergenti. DNA was extracted from the female fly species, then the PCR reaction was amplified by two primers (LITSR, L5.8S) that transcribed a partial internal transcribed spacer (ITS)-1 gene for Leishmania parasite with a length of 320 bp. PCR showed the presence of Leishmania DNA in females of both P. papatasi (10%) and P. sergenti (20%). To determine Leishmania species transmitted by the two previous fly species, the RFLP-PCR technique was performed by the HaeIII enzyme for Leishmania DNA extracted from them. RFLP-PCR showed that P. papatasi females transmitted Leishmania major and P. sergenti females transmitted Leishmania tropica in Refugee camps. It could be concluded that leishmaniasis is widely distributed in Refugee camps due to the presence of its vector.

Keywords: HaeIII enzyme; P. papatasi; P. sergentii; RFLP-PCR; Leishmania