Journal of Biotechnology & Biomaterials
Open Access

Abstract

P.caucasicus and P. mongolensis, subgenus of Paraphlebotomous, are secondary vectors of Zoonotic Cutaneous Leishmaniasis in Iran country. These are two closely related, morphologically similar sand fly species that we cannot find any differentiations between females of them by the standard identifiably keys. This study was carried out in order to verify identification and differentiation of P.caucasicus and P. mongolensis by PCR- restriction enzyme method. Genomic DNA was extracted from 12 specimens of P.caucasicus (male), 3 specimens of P.mongolensis (male) and 9 specimens of Cucasicus Group (female). We selected two primers from the species of sand flies and then ITS2 region of the rDNA gene was amplified by a Polymerase Chain Reaction (PCR). Primers selections were banded on alignment of ITS2 ribosomal DNA by using DNASIS software. By the PCR, 9 specimens of P.caucasicus (male), 2 specimens of P.mongolensis (male) and 6 specimens of Cucasicus Group (female) showed distinguish identical patterns with a visible fragment of about 500 bp in size. In RFLP, we had identical pattern with TasI used as restriction enzymes. For 5 specimens of three species of sand flies isolates, P.caucasicus P. mongolensis and P.papatasi, nucleotide sequencing was achieved, and comparison with other available sequences in GenBank was performed using Blast program. Two of them were the species of P.caucasicus (male), also two of another was P.mongolensis (male) and the fifth was P.papatasi. It is presumed that using PCR with specific primers could be useful for molecular analysis, diagnosis, epidemiological studies and differentiation between P.caucasicus and P. mongolensis.

Biography

Alireza Zahraei is a PhD research Scholar in Banaras Hindu University, Varanasi, India. He was graduated in two courses of Environmental Health and Medical Entomology and Vector Control in BSc and also postgraduate in Medical Entomology and Vector Control. He is a Lecturer in Department of Medical Entomology and vector control, School of Public Health, Tehran University of Medical Sciences, Iran. He has 36 published articles in different international journals and also 47 published in different congress around the world.