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August 20, 2026Scientific ReportsOpen Access

Preliminary evaluation of a qPCR HRM assay for differentiation of Leishmania major and Leishmania tropica

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Authors

YSYousef SharifiARAmirali ReihaniDADonya Akhlaghi

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Overview

Diagnostic study demonstrates accurate differentiation of Leishmania species using real-time PCR high-resolution melting, highlighting its potential for rapid cutaneous leishmaniasis surveillance.

Key Points

  • To evaluate a quantitative real-time PCR coupled with high-resolution melting (qPCR-HRM) assay targeting kinetoplast minicircle DNA (kDNA) for differentiating Leishmania major and Leishmania tropica.
  • Examined clinical samples from 60 patients with suspected cutaneous leishmaniasis in North Khorasan and Razavi Khorasan provinces, Iran, using microscopy and promastigote culture.
  • Developed a qPCR-HRM assay targeting kDNA and validated results against conventional PCR and ITS1-rDNA sequencing.
  • qPCR-HRM differentiated reference strains by distinct melting temperatures (Tm = 84.13 °C for L. major vs 83.27 °C for L. tropica; mean ΔTm ≈ 0.86 °C).
  • Identified 13 molecularly positive samples out of 60, with 100% concordance between qPCR-HRM and ITS1 sequencing identifying all 13 as L. major, comprising nine sequence types and seven haplotypes.

Cite This Study

Sharifi et al. (2026) studied this question.

synapsesocial.com/papers/6a86b5978a91293e6a1ccffdhttps://doi.org/10.1038/s41598-026-66911-5
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  5. 5Identification of Causative Species in Cutaneous Leishmaniasis Patients Using PCR-RFLP2011 · 14 citations