{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T16:17:48Z","timestamp":1778084268839,"version":"3.51.4"},"reference-count":36,"publisher":"American Society of Tropical Medicine and Hygiene","issue":"5","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,5,6]]},"abstract":"<jats:title>ABSTRACT.<\/jats:title>\n                  <jats:p>\n                    Microscopic examination (ME) of dermal scrapings is widely used for biological confirmation of cutaneous leishmaniasis (CL), a major health concern in North Africa. This study aims to evaluate this technique and to assess the contribution of stained smears as the DNA source for polymerase chain reaction (PCR). In total, 227 archived Giemsa-stained smears from CL-suspected patients were examined by a standardized graded microscopy followed by scraping, DNA extraction, and kinetoplast DNA real-time PCR amplification. Examination of two 100-mm\n                    <jats:sup>2<\/jats:sup>\n                    squares for 10 minutes each identified 76 positive slides with varying parasitic loads (1 to &gt;40 amastigotes) and amastigote detection times (range: 1 minute to 18 minutes and 10 seconds). Polymerase chain reaction was more sensitive than ME (99% versus 75.2%), detecting 25 additional positive cases with low estimated parasitic load (median: 2 parasites\/slide, interquartile range: 0.5\u20135.7). This combined approach associates the advantages of microscopy with the increased sensitivity of PCR, being particularly interesting in ME-negative cases with strong clinical suspicion.\n                  <\/jats:p>","DOI":"10.4269\/ajtmh.25-0519","type":"journal-article","created":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T15:01:38Z","timestamp":1774969298000},"page":"870-873","source":"Crossref","is-referenced-by-count":0,"title":["Microscopy and Polymerase Chain Reaction on Giemsa-Stained Smears: A Combined Approach for Cutaneous Leishmaniasis Diagnosis"],"prefix":"10.4269","volume":"114","author":[{"given":"Fida","family":"Maatallah","sequence":"first","affiliation":[{"name":"Laboratory of Epidemiology and Ecology of Parasites, Pasteur Institute of Tunis, Tunis, Tunisia;"},{"name":"Research Laboratory 16-IPT-06, Medical Parasitology, Biotechnology and Biomolecules, Pasteur Institute of Tunis, University of Tunis-El Manar, Tunis, Tunisia;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wafa","family":"Aissi","sequence":"additional","affiliation":[{"name":"National Center for the Promotion of Organ Transplantation, Bab Saadoun, Tunisia;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Olfa","family":"Souissi","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Pasteur Institute of Tunis, Tunis, Tunisia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nada","family":"Boulehmi","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Pasteur Institute of Tunis, Tunis, Tunisia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A\u00efda","family":"Bouratbine","sequence":"additional","affiliation":[{"name":"Research Laboratory 16-IPT-06, Medical Parasitology, Biotechnology and Biomolecules, Pasteur Institute of Tunis, University of Tunis-El Manar, Tunis, Tunisia;"},{"name":"Department of Parasitology, Pasteur Institute of Tunis, Tunis, Tunisia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karim","family":"Aoun","sequence":"additional","affiliation":[{"name":"Laboratory of Epidemiology and Ecology of Parasites, Pasteur Institute of Tunis, Tunis, Tunisia;"},{"name":"Research Laboratory 16-IPT-06, Medical Parasitology, Biotechnology and Biomolecules, Pasteur Institute of Tunis, University of Tunis-El Manar, Tunis, Tunisia;"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2795","reference":[{"key":"b1","series-title":"Lancet Infect Dis","first-page":"581","article-title":"Cutaneous leishmaniasis","volume":"7","author":"Reithinger","year":"2007"},{"key":"b2","series-title":"Parasite","first-page":"14","article-title":"Cutaneous leishmaniasis in North Africa: A review","volume":"21","author":"Aoun","year":"2014"},{"key":"b3","series-title":"Microorganisms","first-page":"1632","article-title":"Laboratory diagnosis of cutaneous and visceral leishmaniasis: Current and future methods","volume":"8","author":"Reim\u00e3o","year":"2020"},{"key":"b4","series-title":"Tunis Med","first-page":"823","article-title":"Epidemiological and clinical criteria for the diagnosis of cutaneous leishmaniasis in Tunisia","volume":"103","author":"Aoun","year":"2025"},{"key":"b5","article-title":"Leishmaniasis.","year":"2025"},{"key":"b6","series-title":"Clin Infect Dis","first-page":"e202","article-title":"Diagnosis and treatment of leishmaniasis: Clinical practice guidelines by the Infectious Diseases Society of America (IDSA) and the American Society of Tropical Medicine and Hygiene (ASTMH)","volume":"63","author":"Aronson","year":"2016"},{"key":"b7","series-title":"Bull Soc Pathol Exot","first-page":"251","article-title":"Contribution of real-time PCR in the diagnosis of cutaneous leishmaniasis: Experience of the Pasteur Institute of Tunis","volume":"113","author":"Aoun","year":"2020"},{"key":"b8","series-title":"Iran J Parasitol","first-page":"1","article-title":"Comparison of three methods for diagnosis of cutaneous leishmaniasis","volume":"5","author":"Pourmohammadi","year":"2010"},{"key":"b9","series-title":"Am J Trop Med Hyg","first-page":"905","article-title":"Press imprint smear: A rapid, simple, and cheap method for the diagnosis of cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis.","volume":"91","author":"Sousa","year":"2014"},{"key":"b10","series-title":"J Clin Microbiol","first-page":"601","article-title":"PCR-based diagnosis of acute and chronic cutaneous leishmaniasis caused by Leishmania (Viannia)","volume":"40","author":"Weigle","year":"2002"},{"key":"b11","series-title":"Front Microbiol","first-page":"1907","article-title":"Analytical performance of four polymerase chain reaction (PCR) and real time PCR (qPCR) assays for the detection of six Leishmania species DNA in Colombia","volume":"8","author":"Le\u00f3n","year":"2017"},{"key":"b12","series-title":"J Clin Microbiol","first-page":"5249","article-title":"Quantification of Leishmania infantum DNA by a real-time PCR assay with high sensitivity","volume":"42","author":"Mary","year":"2004"},{"key":"b13","series-title":"Microorganisms","first-page":"1157","article-title":"Imaging Leishmania major antigens in experimentally infected macrophages and dermal scrapings from cutaneous leishmaniasis lesions in Tunisia","volume":"10","author":"Sa\u00efdi","year":"2022"},{"key":"b14","series-title":"Acta Trop","first-page":"55","article-title":"Clinical diagnosis of cutaneous leishmaniasis: A comparison study between standardized graded direct microscopy and ITS1-PCR of Giemsa-stained smears","volume":"99","author":"Al-Jawabreh","year":"2006"},{"key":"b15","series-title":"Ann Trop Med Parasitol","first-page":"31","article-title":"DNA extraction and amplification of Leishmania from archived, Giemsa-stained slides, for the diagnosis of cutaneous leishmaniasis by PCR","volume":"96","author":"Motazedian","year":"2002"},{"key":"b16","series-title":"Parasitology","first-page":"1891","article-title":"Sensitive diagnosis of cutaneous leishmaniasis by lesion swab sampling coupled to qPCR","volume":"141","author":"Adams","year":"2014"},{"key":"b17","series-title":"Rev Inst Med Trop Sao Paulo","first-page":"e38","article-title":"Efficiency of noninvasive sampling methods (swab) together with polymerase chain reaction (PCR) for diagnosing American tegumentary leishmaniasis","volume":"59","author":"Boni","year":"2017"},{"key":"b18","series-title":"Am J Trop Med Hyg","first-page":"518","article-title":"Diagnosis of cutaneous leishmaniasis: Why punch when you can scrape?","volume":"92","author":"Saab","year":"2015"},{"key":"b1","series-title":"Lancet Infect Dis","first-page":"581","article-title":"Cutaneous leishmaniasis","volume":"7","author":"Reithinger","year":"2007"},{"key":"b2","series-title":"Parasite","first-page":"14","article-title":"Cutaneous leishmaniasis in North Africa: A review","volume":"21","author":"Aoun","year":"2014"},{"key":"b3","series-title":"Microorganisms","first-page":"1632","article-title":"Laboratory diagnosis of cutaneous and visceral leishmaniasis: Current and future methods","volume":"8","author":"Reim\u00e3o","year":"2020"},{"key":"b4","series-title":"Tunis Med","first-page":"823","article-title":"Epidemiological and clinical criteria for the diagnosis of cutaneous leishmaniasis in Tunisia","volume":"103","author":"Aoun","year":"2025"},{"key":"b5","article-title":"Leishmaniasis.","year":"2025"},{"key":"b6","series-title":"Clin Infect Dis","first-page":"e202","article-title":"Diagnosis and treatment of leishmaniasis: Clinical practice guidelines by the Infectious Diseases Society of America (IDSA) and the American Society of Tropical Medicine and Hygiene (ASTMH)","volume":"63","author":"Aronson","year":"2016"},{"key":"b7","series-title":"Bull Soc Pathol Exot","first-page":"251","article-title":"Contribution of real-time PCR in the diagnosis of cutaneous leishmaniasis: Experience of the Pasteur Institute of Tunis","volume":"113","author":"Aoun","year":"2020"},{"key":"b8","series-title":"Iran J Parasitol","first-page":"1","article-title":"Comparison of three methods for diagnosis of cutaneous leishmaniasis","volume":"5","author":"Pourmohammadi","year":"2010"},{"key":"b9","series-title":"Am J Trop Med Hyg","first-page":"905","article-title":"Press imprint smear: A rapid, simple, and cheap method for the diagnosis of cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis.","volume":"91","author":"Sousa","year":"2014"},{"key":"b10","series-title":"J Clin Microbiol","first-page":"601","article-title":"PCR-based diagnosis of acute and chronic cutaneous leishmaniasis caused by Leishmania (Viannia)","volume":"40","author":"Weigle","year":"2002"},{"key":"b11","series-title":"Front Microbiol","first-page":"1907","article-title":"Analytical performance of four polymerase chain reaction (PCR) and real time PCR (qPCR) assays for the detection of six Leishmania species DNA in Colombia","volume":"8","author":"Le\u00f3n","year":"2017"},{"key":"b12","series-title":"J Clin Microbiol","first-page":"5249","article-title":"Quantification of Leishmania infantum DNA by a real-time PCR assay with high sensitivity","volume":"42","author":"Mary","year":"2004"},{"key":"b13","series-title":"Microorganisms","first-page":"1157","article-title":"Imaging Leishmania major antigens in experimentally infected macrophages and dermal scrapings from cutaneous leishmaniasis lesions in Tunisia","volume":"10","author":"Sa\u00efdi","year":"2022"},{"key":"b14","series-title":"Acta Trop","first-page":"55","article-title":"Clinical diagnosis of cutaneous leishmaniasis: A comparison study between standardized graded direct microscopy and ITS1-PCR of Giemsa-stained smears","volume":"99","author":"Al-Jawabreh","year":"2006"},{"key":"b15","series-title":"Ann Trop Med Parasitol","first-page":"31","article-title":"DNA extraction and amplification of Leishmania from archived, Giemsa-stained slides, for the diagnosis of cutaneous leishmaniasis by PCR","volume":"96","author":"Motazedian","year":"2002"},{"key":"b16","series-title":"Parasitology","first-page":"1891","article-title":"Sensitive diagnosis of cutaneous leishmaniasis by lesion swab sampling coupled to qPCR","volume":"141","author":"Adams","year":"2014"},{"key":"b17","series-title":"Rev Inst Med Trop Sao Paulo","first-page":"e38","article-title":"Efficiency of noninvasive sampling methods (swab) together with polymerase chain reaction (PCR) for diagnosing American tegumentary leishmaniasis","volume":"59","author":"Boni","year":"2017"},{"key":"b18","series-title":"Am J Trop Med Hyg","first-page":"518","article-title":"Diagnosis of cutaneous leishmaniasis: Why punch when you can scrape?","volume":"92","author":"Saab","year":"2015"}],"container-title":["The American Journal of Tropical Medicine and Hygiene"],"original-title":[],"link":[{"URL":"https:\/\/www.ajtmh.org\/view\/journals\/tpmd\/114\/5\/article-p870.xml","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.ajtmh.org\/downloadpdf\/journals\/tpmd\/114\/5\/article-p870.xml","content-type":"text\/html","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/www.ajtmh.org\/downloadpdf\/journals\/tpmd\/114\/5\/article-p870.xml","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T15:37:26Z","timestamp":1778081846000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.ajtmh.org\/view\/journals\/tpmd\/114\/5\/article-p870.xml"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,6]]},"references-count":36,"journal-issue":{"issue":"5"},"URL":"https:\/\/doi.org\/10.4269\/ajtmh.25-0519","relation":{},"ISSN":["0002-9637","1476-1645"],"issn-type":[{"value":"0002-9637","type":"print"},{"value":"1476-1645","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,6]]}}}