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Exploring the Potential of Taq Man Quantitative Pcr for the Simulated Diagnosis of Cutaneous and Visceral Leishmaniasis in Clinical Samples in Iran Publisher



Roozbehani M1 ; Tasbihi M2 ; Keyvani H1, 3 ; Mosavizadeh L3 ; Hasanpour H4 ; Askari Z5
Authors
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Authors Affiliations
  1. 1. Vaccine Research Center, Iran University of Medical Sciences, Tehran, Iran
  2. 2. Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran
  3. 3. Department of Virology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
  4. 4. Department of Parasitology and Mycology, School of Allied Medical Sciences, Ilam University of Medical Sciences, Ilam, Iran
  5. 5. Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

Source: Journal of Parasitic Diseases Published:2025


Abstract

Leishmaniasis is a major neglected tropical disease that can lead to fatalities among infected individuals. Clinical identification was based on microscopic examination and parasitological culture performed by trained technicians. The limited accuracy and inconvenience associated with the microscopic analysis may lead to the misdiagnosis and recurrence of leishmaniasis. Consequently, an in-house TaqMan quantitative PCR (qPCR) method using the kinetoplast minicircle DNA (mkDNA) gene was developed simultaneously diagnose cutaneous and visceral leishmaniasis in clinical specimens. A total of 77 skin lesion samples, 10 canine blood samples, aspirates and 65 samples as control were confirmed by microscopy, in vitro cultured promastigotes, and rK39 rapid diagnostic tests. The mkDNA gene was analyzed by qPCR to determine the detection limit, sensitivity, and specificity of the test. These results demonstrated a sensitivity of 96.3% (95 CI 81.03–99.91%), specificity of 100.00% (95 CI 94.04–100.00%), and accuracy of 98.85% (95 CI 93.76–99.97%). The test efficiency ranged from 70 to 97%, with an R2 value of 0.988. The qPCR assay established in this study is a valuable tool for diagnosing cutaneous and visceral leishmaniasis. It is easier to perform than parasitological exanimation because it saves time and reduces the risk of contamination for clinical surveillance and determination of the incidence of leishmaniasis in Iran, an endemic region. This has paved the way for other researchers to explore commercial TaqMan real-time PCR diagnostic kits in Iran. © Indian Society for Parasitology 2025.
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