{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T15:10:19Z","timestamp":1761491419376,"version":"build-2065373602"},"reference-count":45,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2022,6,4]],"date-time":"2022-06-04T00:00:00Z","timestamp":1654300800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004409","name":"Ministry of High Education and Research","doi-asserted-by":"publisher","award":["LR 20IPT06"],"award-info":[{"award-number":["LR 20IPT06"]}],"id":[{"id":"10.13039\/501100004409","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Pasteur Institute of Tunis","award":["LR 20IPT06"],"award-info":[{"award-number":["LR 20IPT06"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Leishmania major cutaneous leishmaniasis (CL) lesions are characterized by an intense process of parasite destruction and antigen processing that could limit microscopic amastigote detection. The aim of our study was to develop a direct immunofluorescence (DIF) assay for in situ visualization of L. major antigens and access its reliability in the routine diagnosis of CL. The developed DIF assay used IgG polyclonal antibodies produced in rabbits by intravenous injections of live L. major metacyclic promastigotes chemically coupled to fluorescein isothiocyanate. Applied to L. major infected RAW macrophages, corresponding macrophage-derived amastigotes and dermal scrapings from CL lesions, the immunofluorescence assay stained specifically Leishmania amastigotes and showed a diffuse Leishmania antigen deposit into cytoplasm of phagocytic cells. Reliability of DIF in CL diagnosis was assessed on 101 methanol-fixed dermal smears from 59 positive and 42 negative CL lesions diagnosed by direct microscopy and\/or kDNA real-time PCR. Sensitivity and specificity of DIF was 98.3% and 100%, respectively, being more sensitive than microscopy (p &lt; 0.001) and as sensitive as ITS1-PCR. ITS1-PCR-RFLP allowed Leishmania species identification in 56 out of the 58 DIF-positive smears, identifying 52 L. major, two L. infantum and two L. tropica cases, which indicates antigenic cross-reactivity between Leishmania species.<\/jats:p>","DOI":"10.3390\/microorganisms10061157","type":"journal-article","created":{"date-parts":[[2022,6,4]],"date-time":"2022-06-04T09:42:32Z","timestamp":1654335752000},"page":"1157","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Imaging Leishmania major Antigens in Experimentally Infected Macrophages and Dermal Scrapings from Cutaneous Leishmaniasis Lesions in Tunisia"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5036-1811","authenticated-orcid":false,"given":"Nasreddine","family":"Sa\u00efdi","sequence":"first","affiliation":[{"name":"Laboratoire de Recherche, Parasitoses M\u00e9dicales, Biotechnologies et Biomol\u00e9cules, LR 20-IPT-06, Institut Pasteur de Tunis, Universit\u00e9 Tunis El-Manar, Tunis 1002, Tunisia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yousr","family":"Gala\u00ef","sequence":"additional","affiliation":[{"name":"Laboratoire de Recherche, Parasitoses M\u00e9dicales, Biotechnologies et Biomol\u00e9cules, LR 20-IPT-06, Institut Pasteur de Tunis, Universit\u00e9 Tunis El-Manar, Tunis 1002, Tunisia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meriem","family":"Ben-Abid","sequence":"additional","affiliation":[{"name":"Laboratoire de Recherche, Parasitoses M\u00e9dicales, Biotechnologies et Biomol\u00e9cules, LR 20-IPT-06, Institut Pasteur de Tunis, Universit\u00e9 Tunis El-Manar, Tunis 1002, Tunisia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thouraya","family":"Boussoffara","sequence":"additional","affiliation":[{"name":"Laboratoire de Recherche, Transmission, Contr\u00f4le et Immunobiologie des Infections, LR 20-IPT-02, Institut Pasteur de Tunis, Universit\u00e9 Tunis El-Manar, Tunis 1002, Tunisia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ines","family":"Ben-Sghaier","sequence":"additional","affiliation":[{"name":"Laboratoire de Recherche, Parasitoses M\u00e9dicales, Biotechnologies et Biomol\u00e9cules, LR 20-IPT-06, Institut Pasteur de Tunis, Universit\u00e9 Tunis El-Manar, Tunis 1002, Tunisia"},{"name":"Service de Parasitologie-Mycologie, Institut Pasteur de Tunis, Tunis 1002, Tunisia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karim","family":"Aoun","sequence":"additional","affiliation":[{"name":"Laboratoire de Recherche, Parasitoses M\u00e9dicales, Biotechnologies et Biomol\u00e9cules, LR 20-IPT-06, Institut Pasteur de Tunis, Universit\u00e9 Tunis El-Manar, Tunis 1002, Tunisia"},{"name":"Service de Parasitologie-Mycologie, Institut Pasteur de Tunis, Tunis 1002, Tunisia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7049-7030","authenticated-orcid":false,"given":"A\u00efda","family":"Bouratbine","sequence":"additional","affiliation":[{"name":"Laboratoire de Recherche, Parasitoses M\u00e9dicales, Biotechnologies et Biomol\u00e9cules, LR 20-IPT-06, Institut Pasteur de Tunis, Universit\u00e9 Tunis El-Manar, Tunis 1002, Tunisia"},{"name":"Service de Parasitologie-Mycologie, Institut Pasteur de Tunis, Tunis 1002, Tunisia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,4]]},"reference":[{"key":"ref_1","unstructured":"(2022, May 27). 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