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

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.