Evaluative Assay of Nuclear and Mitochondrial Genes to Diagnose Leishmania Species in Clinical Specimens

Leishmaniasis as an emerging and reemerging disease is increasing worldwide with high prevalence and new incidence in recent years. For epidemiological investigation and accurate identification of species, three nuclear and mitochondrial genes (ITS-rDNA, Hsp70, and Cyt ) were employed and analyzed f...

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Bibliographic Details
Published in:Iranian journal of public health 2017-10, Vol.46 (10), p.1422-1429
Main Authors: Esmaeili Rastaghi, Ahmad Reza, Spotin, Adel, Khataminezhad, Mohammad Reza, Jafarpour, Mostafa, Alaeenovin, Elnaz, Najafzadeh, Narmin, Samei, Neda, Taleshi, Neda, Mohammadi, Somayeh, Parvizi, Parviz
Format: Article
Language:English
Subjects:
Cutaneous leishmaniasis
Cyt b
Deoxyribonucleic acid
Diagnosis
DNA
Epidemiology
Genes
Hsp70
Hsp70 protein
Identification methods
Iran
ITS-rDNA
Laboratories
Laboratory methods
Leishmania
Leishmania species
Lesions
Mitochondria
Parasites
Parasitic diseases
Patients
Polymerase chain reaction
Protozoa
Sensitivity analysis
Short Communication
Species
Vector-borne diseases
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Summary:Leishmaniasis as an emerging and reemerging disease is increasing worldwide with high prevalence and new incidence in recent years. For epidemiological investigation and accurate identification of species, three nuclear and mitochondrial genes (ITS-rDNA, Hsp70, and Cyt ) were employed and analyzed from clinical samples in three important Zoonotic Cutaneous Leishmaniasis (ZCL) foci of Iran. In this cross-sectional/descriptive study conducted in 2014-15, serous smears of lesions were directly prepared from suspected patients of ZCL in Turkmen in northeast, Abarkouh in center and Shush district in southwest of Iran. They were directly prepared from suspected patients and DNA was extracted. Two nuclear genes of ITS-rDNA, Hsp70 and one mitochondrial gene of Cyt within parasites were amplified. RFLP was performed on PCR-positive samples. PCR products were sequenced, aligned and edited with sequencher 4.1.4 and phylogenic analyses performed using MEGA 5.05 software. Overall, 203 out of 360 clinical samples from suspected patients were positive using routine laboratory methods and 231 samples were positive by molecular techniques. , and were firmly identified by employing different molecular genes and phylogenic analyses. By combining different molecular genes, parasites were identified accurately. The sensitivity and specificity three genes were evaluated and had more advantages to compare routine laboratory methods. ITS-rDNA gene is more appropriate for firm identification of species.
ISSN:2251-6085
2251-6093