HRM–PCR is an accurate and sensitive technique for the diagnosis of cutaneous leishmaniasis as compared with conventional PCR

Abstract

Background

Cutaneous leishmaniasis (CL) is considered one of the main health problems in Iran. Therefore, it is required for control and therapeutic purposes, an accurate and fast tool for the diagnosis and identification of Leishmania species.

Methods

In the present study, three techniques, including microscopic examination, conventional PCR, and high-resolution melting (HRM)–PCR, have been evaluated, to find the most accurate and rapid test. In total, 105 skin scraping smears were taken from suspected dermal lesions of patients belonging to two known endemic CL areas, Gonbad and Bam districts, in Iran. Subsequently, the specimens were analyzed with microscopic, conventional PCR, and HRM–PCR techniques.

Results

Most positive samples (89.5%) were observed using HRM–PCR, and among the three techniques, HRM–PCR was the most sensitive (89%, 95% CI 81–94) technique. Microscopic examination test had the lowest sensitivity (57%, 95% CI 47–66%). The highest agreement among positive samples was observed between HRM–PCR and conventional PCR tests.

Discussion

Our results showed that the HRM–PCR technique is the most accurate and sensitive test for recognizing CL, and also a valuable alternative test for conventional PCR to detect various species.

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Acknowledgements

The authors would like to thank all participants who contributed to our study.

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Affiliations

Authors

Contributions

MF and RV designed the study, supervised the laboratory work and finalized and revised the manuscript. SNE collected some skin smears of CL patients and review the manuscript. MD performed the microscopic examination of the skin samples. SA and NM carried out the DNA extraction and PCR assays, analysis and wrote the first draft of the manuscript. HZH performed data processing and statistical analysis. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Mahdi Fakhar.

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Conflict of interests

No competing interests were disclosed.

Ethics approval and consent to participate

This study was approved by the ethical committee at Mazandaran University of Medical Sciences (IR.MAZUMS.REC.1395.1102). Participants gave their informed written consent for skin scrapings of their sores to be taken for research purposes.

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Asfaram, S., Fakhar, M., Mirani, N. et al. HRM–PCR is an accurate and sensitive technique for the diagnosis of cutaneous leishmaniasis as compared with conventional PCR. Acta Parasit. 65, 310–316 (2020). https://doi.org/10.2478/s11686-019-00154-5

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Keywords

  • Cutaneous leishmaniasis
  • Leishmania spp.
  • Conventional PCR
  • HRM–PCR