Article

PCR-based detection of Leishmania major kDNA within naturally infected Phlebotomus papatasi in southern Iran

Department of Medical Entomology, Research Centre for Health Sciences, School of Health and Nutrition, Shiraz University of Medical Sciences, P.O. Box 71645-111, Shiraz, Iran.
Transactions of the Royal Society of Tropical Medicine and Hygiene (Impact Factor: 1.93). 06/2010; 104(6):440-2. DOI: 10.1016/j.trstmh.2010.01.008
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ABSTRACT The annual incidence of cutaneous leishmaniasis (CL) in Iran rose by 43% over a five year period, from 2002 to 2006; most of these cases were caused by Leishmania major. Two complementary standard and nested polymerase chain reactions (PCRs) were used to detect parasites within their natural vector, Phlebotomus papatasi. Twelve different sand fly species were morphologically identified. The most abundant species (31.3%) was P. papatasi. Leptomonads were found in nine (2.4%) phlebotomines. Twenty (5.3%) sand fly species were found positive for Leishmania-genus DNA using standard PCR. The infection rate of this species was 5% and 7% by microscopic and molecular methods, respectively.

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    • "In Iran different PCR-based methods including RAPD-PCR and conventional PCRs targeting various genomic DNA fragments kDNA, have been employed for identification of Leishmania parasite species in clinical samples from patients, reservoir hosts, and phlebotomine sand flies (Hajjaran et al., 2013c; Azizi et al., 2010). This methodology has provided some valuable information about diversity of Leishmania parasite species in Iran, e.g. "
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    ABSTRACT: The identity of Iranian Leishmania species has been resolved to some extent by some genetic markers. In this study, based on N-Acetylglucosamine-1-Phosphate Transferase (nagt) gene, we further elucidated the identity and phylogeny of the prevalent species in this country. DNAs of 121 isolates belonging to cutaneous leishmainasis (CL) patients, canine visceral leishmaniasis (CVL) cases, and Rhombomys opimus rodents were amplified by targeting a partial sequence of nagt gene. All the amplicons were analyzed with restriction fragment length polymorphism (RFLP) using Acc1 enzyme, and 49 amplicons representing different reservoir hosts were sequenced and aligned with similar sequences from Genbank database. The RFLP analysis revealed that 41 CL patients were infected L. tropica and 36 with L. major. Among 10 CVL isolates, 6 were identified as L. infantum and 4 as L. tropica. Amongst 34 rodents' isolates, 11 and 23 isolates exhibited patterns similar to those of L. major, and L. tropica/ L. turanica, respectively. The sequencing results from all CL patients, CVL cases, and 4 reservoir rodents were in agreement with RFLP analysis and showed 99-100% homologies with the registered species of L. major, L. tropica, and L. infantum from Turkey, Tunisia, Iraq and Israel. Of the 7 rodent isolates exhibiting RFLP patterns similar to L. tropica/L. turanica, 3 exhibited the highest homologies (99-100%) with L. turanica and 4 with L. gerbilli. The 49 nagt DNA sequences were grouped into five clusters representing L. major, L. tropica, L. infantum, L. turanica and L. gerbilli species, encompassing 19 haplotypes. No correlation was observed between intraspecies divergence and geographic distribution of haplotypes. The L. tropica haplotypes exhibited more homologies with those of L. infantum than L. major (97.2% vs. 96.9%), a probable indication to the potential ability of L. tropica to visceralize. Characterization of Iranian Leishmania isolates using nagt gene allowed unambiguous identification of five prevalent species with a high-resolution phylogeny.
    Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 06/2014; 26. DOI:10.1016/j.meegid.2014.05.026 · 3.26 Impact Factor
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    • "The true number may be 4 times more than this figure and in fact this disease is considered one of the most important parasitic diseases after malaria. The most likely cause for this upward trend is preponderance in human-sand fly contact, which is probably the product of the development of irrigation schemes and the spread of human populations into the habitats of the vector, Phlebotomus papatasi Scopoli, and the rodents that act as reservoir hosts [4] [5] [6] [7] [8] [9] [10] [11] [12] . CL can be seen in zoonotic or anthroponotic forms. "
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    ABSTRACT: Objective: To investigate the prevalence of cutaneous leishmaniasis in important endemic focus of Kashan district, central of Iran during 2007-2008. Methods: This was a cross-sectional study and 5 098 individuals were selected from thirteen rural and urban districts of Kashan city. Information of positive cases including age, sex, job, number and sites of ulcers or scars, date and place of the ulcer, and results of clinical examination and laboratory tests were recorded. Diagnoses of affected cases were based on clinical examination and microscopic observation of the ulcer parasite. The results were analyzed with statistical Chi-square test. Results: An infection prevalence rate of 6.4% was obtained among 5 098 individuals. Regarding to 326 affected cases, 103 (2.0%) and 223 (4.4%) persons were observed with active lesion and scars, respectively. The highest frequency of active ulcer rate (23.3%) was associated with age group of 20-29 years old, while the lowest rate was related to age group of 0-9 years old with 7.8% infection. About 49.5% of the infected cases were under 30 years old. This study showed 64.1% of cases with one and the rest of them with two or more ulcers. Hands (46.6%) were the most affected parts of body. Conclusions: There is a progressive increasing of cutaneous leishmaniasis in Kashan district and this is a warning to local health workers to provide prevention as well as control program of the disease.
    08/2012; 2(4). DOI:10.1016/S2222-1808(12)60057-7
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    • "Cortes et al. (2006) applied kDNA as a molecular marker to analyze L. infantum diversity in Portugal. Also, Aziz et al. (2010) used PCR-based technique to detection of L. major kDNA within naturally infected Phlebotomus papatasi in southern Iran. Recently, Fraga et al. (2010) studied phylogeny of Leishmania species based on the heat-shock protein 70 gene. "
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    ABSTRACT: Fifty samples of skin ulcers were collected from the western region of Saudi Arabia Kingdom (Al Baha and Al Qasim) to study genotypic characterization of Cutaneous Leishmaniasis in this area. Thirty-six samples were recorded as Leishmania isolates. The same isolates were subsequently tested with fingerprinting with single arbitrary primers. The primers used derived from the core sequence of the phage M13, and the repeat sequences (GTG)5 and (GACA)4. The 36 isolates were all identified as Leishmania major (n = 25 isolates) or Leishmania tropica (n = 11 isolates). All produced polymorphic patterns, which were grouped depending on the species they belonged to, next to the relevant well-characterized strains of the same species. Within the L. major and L. tropica group the subgroupings formed were mainly related to the geographical origin of the strains. A nested polymerase chain reaction-based schizodeme method for identifying Leishmania kinetoplast minicircle classes was used as a diagnostic tool for L. major and L. tropica.
    Vector borne and zoonotic diseases (Larchmont, N.Y.) 03/2011; 11(7):807-13. DOI:10.1089/vbz.2010.0213 · 2.53 Impact Factor
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