{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T20:22:15Z","timestamp":1787775735924,"version":"build-2784847793"},"reference-count":27,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"1","license":[{"start":{"date-parts":[[2026,2,1]],"date-time":"2026-02-01T00:00:00Z","timestamp":1769904000000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":[],"abstract":"\n Background:<\/jats:title>\n \n The diagnosis of cutaneous\n leishmaniasis<\/jats:italic>\n (CL) is typically based on microscopic examination of Giemsa-stained smear. However, this method can lead to false negative and false positive results. Therefore, the study aimed to identify\n L. major<\/jats:italic>\n and\n L<\/jats:italic>\n .\n tropica<\/jats:italic>\n species by investigating microscopically negative smears suspected of CL using the multiplex nested polymerase chain reaction (PCR) method.\n <\/jats:p>\n <\/jats:sec>\n \n Materials and Methods:<\/jats:title>\n \n This study was conducted on 227 microscopically negative specimens from individuals suspected of having CL referred to the CL Treatment and Diagnosis Center in Gonbad-e Kavus Health Center, Golestan Province, northeastern Iran. Multiplex nested PCR technique was utilized to detect\n L. tropica<\/jats:italic>\n and\n L. major<\/jats:italic>\n under investigation based on\n ITS1<\/jats:italic>\n sequence.\n <\/jats:p>\n <\/jats:sec>\n \n Results:<\/jats:title>\n \n 2.64% (6\/227) of specimens were detected as positive for\n Leishmania<\/jats:italic>\n species, all of which were identified as\n L<\/jats:italic>\n .\n major<\/jats:italic>\n .\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion:<\/jats:title>\n The multiplex nested PCR technique demonstrated that microscopic examination of the smears is linked to false negative in diagnosing CL. Consequently, it is recommended to utilize the multiplex nested PCR method as either a primary diagnostic or a confirmatory approach for cases that are microscopically negative.<\/jats:p>\n <\/jats:sec>","DOI":"10.4103\/abr.abr_539_24","type":"journal-article","created":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T09:00:13Z","timestamp":1772182813000},"update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":0,"title":["Molecular Detection of Leishmania Species on Microscopically Negative Smears in Gonbad-e-Kavus, Northeastern Iran"],"prefix":"10.4103","volume":"15","author":[{"given":"Salehe","family":"Ostovan","sequence":"first","affiliation":[{"name":"Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fatemeh","family":"Livani","sequence":"additional","affiliation":[{"name":"Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Faramarz","family":"Koohsar","sequence":"additional","affiliation":[{"name":"Laboratory Sciences Research Center, Golestan University of Medical Sciences, Gorgan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roghiyeh","family":"Faridnia","sequence":"additional","affiliation":[{"name":"Laboratory Sciences Research Center, Golestan University of Medical Sciences, Gorgan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sima","family":"Besharat","sequence":"additional","affiliation":[{"name":"Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ganesh","family":"Yadagiri","sequence":"additional","affiliation":[{"name":"Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fatemeh","family":"Mesgariyan","sequence":"additional","affiliation":[{"name":"Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammad-Ali","family":"Mohaghegh","sequence":"additional","affiliation":[{"name":"Health Sciences Research Center, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hamed","family":"Kalani","sequence":"additional","affiliation":[{"name":"Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"276","published-online":{"date-parts":[[2026,2]]},"reference":[{"key":"R1-20260302","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1016\/S1473-3099(07)70209-8","article-title":"Cutaneous leishmaniasis","volume":"7","author":"Reithinger","year":"2007","journal-title":"Lancet Infect Dis"},{"key":"R2-20260302","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1007\/s40257-022-00726-8","article-title":"Cutaneous leishmaniasis: A 2022 updated narrative review into diagnosis and management developments","volume":"23","author":"de Vries","year":"2022","journal-title":"Am J Clin Dermatol"},{"key":"R3-20260302","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1111\/j.1529-8019.2009.01272.x","article-title":"Cutaneous and mucocutaneous leishmaniasis","volume":"22","author":"David","year":"2009","journal-title":"Dermatol Ther"},{"key":"R4-20260302","doi-asserted-by":"crossref","first-page":"4103","DOI":"10.1128\/IAI.69.6.4103-4108.2001","article-title":"Protection against cutaneous leishmaniasis induced by recombinant antigens in murine and nonhuman primate models of the human disease","volume":"69","author":"Campos-Neto","year":"2001","journal-title":"Infect Immun"},{"key":"R5-20260302","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1007\/s00403-025-04064-7","article-title":"A twelve-year study of positive cutaneous leishmaniasis cases in Northeastern Iran: A cross-sectional study from 2011 to 2022","volume":"317","author":"Boroumand","year":"2025","journal-title":"Arch Dermatol Res"},{"key":"R6-20260302","first-page":"148","article-title":"Dynamic relations between incidence of zoonotic cutaneous leishmaniasis and climatic factors in Golestan Province, Iran","volume":"9","author":"Shirzadi","year":"2015","journal-title":"J Arthropod Borne Dis"},{"key":"R7-20260302","first-page":"21","article-title":"Molecular detection of Leishmania major in the vectors and reservoir hosts of cutaneous leishmaniasis in Kalaleh District, Golestan Province, Iran","volume":"2","author":"Rassi","year":"2008","journal-title":"J Arthropod Borne Dis"},{"key":"R8-20260302","first-page":"337","article-title":"Molecular identification of Leishmania species using samples obtained from negative stained smears","volume":"8","author":"Mohaghegh","year":"2013","journal-title":"Iran J Parasitol"},{"key":"R9-20260302","doi-asserted-by":"crossref","first-page":"810","DOI":"10.26719\/emhj.23.105","article-title":"Integration and evaluation of cutaneous leishmaniasis laboratory diagnosis in the primary health care laboratory network","volume":"29","author":"Zeinali","year":"2023","journal-title":"East Mediterr Health J"},{"key":"R10-20260302","doi-asserted-by":"crossref","first-page":"610","DOI":"10.3390\/microorganisms9030610","article-title":"Loop-mediated isothermal amplification allows rapid, simple and accurate molecular diagnosis of human cutaneous and visceral leishmaniasis caused by leishmania infantum when compared to PCR","volume":"9","author":"Ibarra-Meneses","year":"2021","journal-title":"Microorganisms"},{"key":"R11-20260302","doi-asserted-by":"crossref","first-page":"e0008903","DOI":"10.1371\/journal.pntd.0008903","article-title":"Evaluation of conventional and four real-time PCR methods for the detection of Leishmania on field-collected samples in Ethiopia","volume":"15","author":"Merdekios","year":"2021","journal-title":"PLoS Negl Trop Dis"},{"issue":"Suppl 1","key":"R12-20260302","doi-asserted-by":"crossref","first-page":"S87","DOI":"10.1016\/S0035-9203(02)90057-X","article-title":"Distinguishing Leishmania tropica and Leishmania major in the Middle East using the polymerase chain reaction with kinetoplast DNA-specific primers","volume":"96","author":"Anders","year":"2002","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"R13-20260302","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1067\/mjd.2003.492","article-title":"Value of diagnostic techniques for cutaneous leishmaniasis","volume":"49","author":"Faber","year":"2003","journal-title":"J Am Acad Dermatol"},{"key":"R14-20260302","doi-asserted-by":"crossref","first-page":"762","DOI":"10.1016\/j.trstmh.2005.06.002","article-title":"Usefulness of PCR in the diagnosis of cutaneous leishmaniasis in Tunisia","volume":"99","author":"Chargui","year":"2005","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"R15-20260302","doi-asserted-by":"crossref","first-page":"2246","DOI":"10.1128\/jcm.32.9.2246-2252.1994","article-title":"Diagnosis of cutaneous leishmaniasis and species discrimination of parasites by PCR and hybridization","volume":"32","author":"Rodr\u00edguez","year":"1994","journal-title":"J Clin Microbiol"},{"key":"R16-20260302","first-page":"54","article-title":"Diagnosis and characterization of Leishmania species in giemsa-stained slides by PCR-RFLP","volume":"37","author":"Kazemi-Rad","year":"2008","journal-title":"Iran J Public Health"},{"key":"R17-20260302","doi-asserted-by":"crossref","first-page":"552","DOI":"10.1016\/j.exppara.2010.06.003","article-title":"Leishmania species: Detection and identification by nested PCR assay from skin samples of rodent reservoirs","volume":"126","author":"Akhavan","year":"2010","journal-title":"Exp Parasitol"},{"key":"R18-20260302","doi-asserted-by":"crossref","first-page":"e0185606","DOI":"10.1371\/journal.pone.0185606","article-title":"Real-time PCR in detection and quantitation of Leishmania donovani for the diagnosis of Visceral Leishmaniasis patients and the monitoring of their response to treatment","volume":"12","author":"Hossain","year":"2017","journal-title":"PLoS One"},{"key":"R19-20260302","doi-asserted-by":"crossref","first-page":"1435","DOI":"10.1128\/JCM.44.4.1435-1439.2006","article-title":"Comparison of PCR assays for diagnosis of cutaneous leishmaniasis","volume":"44","author":"Bensoussan","year":"2006","journal-title":"J Clin Microbiol"},{"key":"R20-20260302","doi-asserted-by":"crossref","first-page":"e0335422","DOI":"10.1128\/spectrum.03354-22","article-title":"Molecular diagnosis of leishmaniasis in Spain: Development and validation of ready-to-use gel-form nested and real-time PCRs to detect Leishmania spp","volume":"11","author":"Chicharro","year":"2023","journal-title":"Microbiol Spectr"},{"key":"R21-20260302","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1186\/1471-2334-12-191","article-title":"Polymorphism-specific PCR enhances the diagnostic performance of American tegumentary leishmaniasis and allows the rapid identification of Leishmania species from Argentina","volume":"12","author":"Marco","year":"2012","journal-title":"BMC Infect Dis"},{"key":"R22-20260302","doi-asserted-by":"crossref","first-page":"1819","DOI":"10.1128\/JCM.37.6.1819-1823.1999","article-title":"Use of PCR in diagnosis of human american tegumentary leishmaniasis in Rio de Janeiro, Brazil","volume":"37","author":"Pirmez","year":"1999","journal-title":"J Clin Microbiol"},{"key":"R23-20260302","doi-asserted-by":"crossref","first-page":"3768","DOI":"10.1128\/JCM.38.10.3768-3773.2000","article-title":"Diagnosis of cutaneous leishmaniasis in Colombia: The sampling site within lesions influences the sensitivity of parasitologic diagnosis","volume":"38","author":"Rami\u0301rez","year":"2000","journal-title":"J Clin Microbiol"},{"key":"R24-20260302","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1159\/000063150","article-title":"Polymerase chain reaction for diagnosis of cutaneous leishmaniasis in histologically positive, suspicious and negative skin biopsies","volume":"205","author":"Safaei","year":"2002","journal-title":"Dermatol"},{"key":"R25-20260302","doi-asserted-by":"crossref","first-page":"756","DOI":"10.1080\/0036554021000026930","article-title":"Differentiation of old and new world Leishmania species at complex and species levels by PCR","volume":"34","author":"Mahboudi","year":"2002","journal-title":"Scand J Infect Dis"},{"key":"R26-20260302","doi-asserted-by":"crossref","first-page":"229","DOI":"10.3347\/kjp.2018.56.3.229","article-title":"Diagnosis, treatment and clinical features of cutaneous Leishmaniasis in Saudi Arabia","volume":"56","author":"Hawash","year":"2018","journal-title":"Korean J Parasitol"},{"key":"R27-20260302","doi-asserted-by":"crossref","first-page":"990","DOI":"10.3390\/microorganisms10050990","article-title":"Development of a highly sensitive nested PCR and its application for the diagnosis of cutaneous leishmaniasis in Sri Lanka","volume":"10","author":"De Silva","year":"2022","journal-title":"Microorganisms"}],"container-title":["Advanced Biomedical Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.ovid.com\/10.4103\/abr.abr_539_24","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T19:40:53Z","timestamp":1787773253000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.ovid.com\/10.4103\/abr.abr_539_24"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2]]},"references-count":27,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026]]}},"URL":"https:\/\/doi.org\/10.4103\/abr.abr_539_24","relation":{},"ISSN":["2277-9175"],"issn-type":[{"value":"2277-9175","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2]]},"assertion":[{"value":"2024-10-28","name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-01","name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}}]}}