{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,19]],"date-time":"2026-07-19T04:05:53Z","timestamp":1784433953187,"version":"3.55.0"},"reference-count":36,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2024,9,4]],"date-time":"2024-09-04T00:00:00Z","timestamp":1725408000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001411","name":"Indian Council of Medical Research (ICMR, India)","doi-asserted-by":"publisher","award":["6\/9-7(228)2020-ECDII"],"award-info":[{"award-number":["6\/9-7(228)2020-ECDII"]}],"id":[{"id":"10.13039\/501100001411","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>The accurate diagnosis and identification of Leishmania species are crucial for the therapeutic selection and effective treatment of leishmaniasis. This study aims to develop and evaluate the use of high-resolution melting curve analysis (HRM)-PCR for Leishmania species identification causing visceral leishmaniasis (VL), post-kala-azar dermal leishmaniasis (PKDL) and cutaneous leishmaniasis (CL) in the Indian subcontinent. Two multi-copy targets (ITS-1 and 7SL-RNA genes) were selected, and an HRM-PCR assay was established using L. donovani, L. major, and L. tropica standard strain DNA. The assay was applied on 93 clinical samples with confirmed Leishmania infection, including VL (n = 30), PKDL (n = 50), and CL (n = 13) cases. The ITS-1 HRM-PCR assay detected as little as 0.01 pg of template DNA for L. major and up to 0.1 pg for L. donovani and L. tropica. The detection limit for the 7SL-RNA HRM-PCR was 1 pg for L. major and 10 pg for L. donovani and L. tropica. The ITS-1 HRM-PCR identified 68 out of 93 (73.11%) leishmaniasis cases, whereas 7SL-RNA HRM-PCR could only detect 18 out of 93 (19.35%) cases. A significant correlation was observed between the kDNA-based low Ct values and ITS-1 HRM-PCR positivity in the VL (p = 0.007), PKDL (p = 0.0002), and CL (p = 0.03) samples. The ITS-1 HRM-PCR assay could identify Leishmania spp. causing different clinical forms of leishmaniasis in the Indian subcontinent, providing rapid and accurate results that can guide clinical management and treatment decisions.<\/jats:p>","DOI":"10.3390\/pathogens13090759","type":"journal-article","created":{"date-parts":[[2024,9,4]],"date-time":"2024-09-04T04:28:28Z","timestamp":1725424108000},"page":"759","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Assessment of High-Resolution Melting Curve Analysis for Leishmania spp. Detection in Different Clinical Manifestations of Leishmaniasis in India"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0554-0017","authenticated-orcid":false,"given":"Mudsser","family":"Azam","sequence":"first","affiliation":[{"name":"ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saurabh","family":"Singh","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Venereology, Leprology, All India Institute of Medical Sciences, Jodhpur 342005, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ratan","family":"Gupta","sequence":"additional","affiliation":[{"name":"VMMC and Safdarjung Hospital, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mayank","family":"Mayank","sequence":"additional","affiliation":[{"name":"ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7213-3924","authenticated-orcid":false,"given":"Sushruta","family":"Kathuria","sequence":"additional","affiliation":[{"name":"VMMC and Safdarjung Hospital, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shruti","family":"Sharma","sequence":"additional","affiliation":[{"name":"ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"V.","family":"Ramesh","sequence":"additional","affiliation":[{"name":"Department of Dermatology and STD, ESIC Medical College & Hospital Faridabad, Faridabad 121012, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8094-4703","authenticated-orcid":false,"given":"Ruchi","family":"Singh","sequence":"additional","affiliation":[{"name":"ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,9,4]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2024, May 10). 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