Real-time PCR using high-resolution melting analysis technology for diagnosis of Leishmania and determination of types of clinical samples

Authors: SERHAT SİREKBASAN, ERDAL POLAT

Abstract: Background/aim: Leishmaniasis is a disease group carried by Phlebotomus and Lutzomyia sand flies infected with Leishmania and is mostly observed in rural areas. In this study, using high-resolution melting analysis (HRMA), we aim to identify the active types of leishmaniasis, which are inadequately identified by classical methods. Materials and methods: Samples of 85 patients were examined in the study. Six of the patients were suspected of having visceral leishmaniasis (VL) and 79 cutaneous leishmaniasis (CL). The slides prepared from the samples were stained with Giemsa stain and examined under a light microscope. The results were compared with those determined in real-time PCR. When the real-time PCR result was positive for Leishmania , we determined the type by HRMA. Results: Among 85 Leishmania amastigote samples, 28 (32.9%) of them were detected and accepted as positive by microscopic examination. On the other hand, 25 (29.4%) of the 85 samples were found as positive using real-time PCR. In addition, when 25 Leishmania -positive samples were examined by HRMA, the results showed that 21 (84%) were L. tropica , 3 (12%) were L. major , and 1 (4%) was L. infantum . Conclusion: Based on our findings, we conclude that using real-time PCR and HRMA is useful in determining the cause of the disease in leishmaniasis.

Keywords: Genotyping, high-resolution melting analysis, Leishmania , leishmaniasis, real-time PCR

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