{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,20]],"date-time":"2026-05-20T20:33:15Z","timestamp":1779309195057,"version":"3.51.4"},"reference-count":32,"publisher":"American Society of Tropical Medicine and Hygiene","issue":"6","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,12,3]]},"abstract":"<jats:title>ABSTRACT.<\/jats:title>\n                  <jats:p>\n                    Accurate\n                    <jats:italic>Leishmania<\/jats:italic>\n                    species identification is crucial for epidemiological aspects and disease management. This study aimed to detect and identify the main causative species of human cutaneous and visceral leishmaniasis in Iran using nested polymerase chain reaction (PCR) based on size polymorphism of the ribosomal DNA (rDNA) intergenic region. In total, 180 clinical samples, including 156 confirmed positive cases, 24 confirmed negative cases, and 33 suspected cases that tested negative by microscopy, were collected from skin lesions, bone marrow, and lymph nodes across five provinces in Iran. DNA was extracted from the samples, and the nested PCR-amplified fragments were differentiated with agarose gel electrophoresis. The nested PCR assay successfully identified\n                    <jats:italic>Leishmania major<\/jats:italic>\n                    ,\n                    <jats:italic>Leishmania tropica<\/jats:italic>\n                    , and\n                    <jats:italic>Leishmania infantum<\/jats:italic>\n                    with high accuracy. Among 156 microscopy-positive samples,\n                    <jats:italic>L. major<\/jats:italic>\n                    was detected in 84 cases,\n                    <jats:italic>L. tropica<\/jats:italic>\n                    was detected in 64 cases, and\n                    <jats:italic>L. infantum<\/jats:italic>\n                    was detected in 8 cases. The results obtained were fully consistent with those from a commercial real-time PCR diagnostic kit. Phylogenetic analysis supported the differentiation of\n                    <jats:italic>Leishmania<\/jats:italic>\n                    species. However, among 33 suspected cases, 14 samples were negative by all three methods (microscopy, commercial PCR kit, and nested PCR), 7 samples tested positive by both molecular methods, 8 samples were positive only with the commercial PCR kit, and 4 samples were positive exclusively by nested PCR. The findings indicate that the nested PCR assay targeting size polymorphisms in the rDNA region is a cost-effective method for detection and identification of\n                    <jats:italic>Leishmania<\/jats:italic>\n                    species in clinical specimens, particularly when microscopy results are inconclusive.\n                  <\/jats:p>","DOI":"10.4269\/ajtmh.24-0843","type":"journal-article","created":{"date-parts":[[2025,10,7]],"date-time":"2025-10-07T15:04:53Z","timestamp":1759849493000},"page":"1220-1225","source":"Crossref","is-referenced-by-count":1,"title":["Detection and Identification of Leishmania major, Leishmania tropica, and Leishmania infantum in Clinical Samples Based on Size Polymorphism of Partially Amplified Ribosomal DNA"],"prefix":"10.4269","volume":"113","author":[{"given":"Reza","family":"Mohammadi Manesh","sequence":"first","affiliation":[{"name":"Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, Iran;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Somayeh","family":"Mousavi","sequence":"additional","affiliation":[{"name":"Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Parisa","family":"Mousavi","sequence":"additional","affiliation":[{"name":"Skin Diseases and Leishmaniasis Research Center, Isfahan University of Medical Sciences, Isfahan, Iran;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Azadeh","family":"Zolfaghari","sequence":"additional","affiliation":[{"name":"Skin Diseases and Leishmaniasis Research Center, Isfahan University of Medical Sciences, Isfahan, Iran;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zabihollah","family":"Zarei","sequence":"additional","affiliation":[{"name":"Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Iraj","family":"Sharifi","sequence":"additional","affiliation":[{"name":"Leishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, Iran;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hossein","family":"Zarrinfar","sequence":"additional","affiliation":[{"name":"Department of Medical Parasitology and Mycology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seyed Hossein","family":"Hejazi","sequence":"additional","affiliation":[{"name":"Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Behrooz","family":"Ataei","sequence":"additional","affiliation":[{"name":"Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, Iran;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mehdi","family":"Mohebali","sequence":"additional","affiliation":[{"name":"Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hossein","family":"Mirhendi","sequence":"additional","affiliation":[{"name":"Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, Iran;"},{"name":"Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran;"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2795","reference":[{"key":"b1","series-title":"Global Leishmaniasis Surveillance: 2019\u20132020, a Baseline for the 2030 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