Nested Polymerase Chain Reaction and Sequence-Based Detection of Leishmania Infection of Sand Flies in Recently Emerged Endemic Focus of Zoonotic Cutaneous Leishmaniasis, Southern Iran
Paper ID : 1160-3IICE
Authors:
Abdollah Badzohreh *
Tehran university of medical sciences, school of public health, department of medical entomology and vector control
Abstract:
Introduction: Geographical distribution of zoonotic cutaneous leishmaniasis (ZCL) has continuously been extended in recent years in Iran. The Beiza District is one of the newly emerged endemic foci of ZCL in southern Iran. The incidence of clinical ZCL cases in this district, was reported as 16.23 and 12.65/1000 in two recent years. The main aim of the present study was to detect the vector(s) of ZCL in this area.

Materials and Methods: To detect the fauna and vectors of ZCL in this study, sand flies were caught using sticky papers from 10 villages. Collection of sand flies was carried out twice a month from April to October. Sixty sticky papers were installed per night at each sampling station such as rural homes and gerbil burrows. Traps were installed after sunset and collected before the next day sunrise. Seventy randomly selected female sand flies out of 730 were molecularly investigated for Leishmania infection using species-specific nested polymerase chain reaction (PCR) assay. The kDNA Leishmania parasites amplification was carried out in two rounds using the primers of CSB1XR and CSB2XF for the first round and LiR and 13Z for the second round.

Results: A total of 2543 sand flies comprised of 730 (28.7%) females and 1813 (71.3%) males were caught. The fauna was identified as 10 species include five Phlebotomus spp. (P. papatasi, P. sergenti, P. tobbi, P. salehi and P. caucasicus) and five Sergentomyia spp. (S. theodori, S. clydei, S. dentate, S. baghdadis and S. squamipleuris). P. papatasi was the most dominant species both indoors and outdoors (37.55% and 16.35 %, respectively). L. major was detected in 5 out of 48 investigated Phlebotomus papatasi (10.41%). Sequence-based characterization was carried out to confirm the PCR findings. The positive samples were shown to have 75-88% similarity with L. major sequences in GenBank.

Conclusion: This is the first report of sand flies from Beiza district. According to the findings of the present study, similar to the other foci of ZCL in Iran, P. papatasi is the proven and primary vector of CL. This study could be drawn upon for future strategy planning in this newly emerged endemic focus.
Keywords:
Leishmaniasis, PCR, Sand flies, Phlebotomus papatasi, Leishmania major, Iran
Status : Paper Accepted (Oral Presentation)