{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T00:11:29Z","timestamp":1790295089653,"version":"4.1.0"},"reference-count":37,"publisher":"Public Library of Science (PLoS)","issue":"1","license":[{"start":{"date-parts":[[2021,1,21]],"date-time":"2021-01-21T00:00:00Z","timestamp":1611187200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006507","name":"Funda\u00e7\u00e3o Oswaldo Cruz","doi-asserted-by":"publisher","award":["VPPCB-008-FIO-18-2-14"],"award-info":[{"award-number":["VPPCB-008-FIO-18-2-14"]}],"id":[{"id":"10.13039\/501100006507","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:sec id=\"sec001\">\n<jats:title>Background<\/jats:title>\n<jats:p>Cutaneous leishmaniasis (CL) is an infectious vector-borne disease caused by protozoa of the <jats:italic>Leishmania<\/jats:italic> genus that affects humans and animals. The distribution of parasites in the lesion is not uniform, and there are divergences in the literature about the choice of the better sampling site for diagnosis\u2013inner or outer edge of the ulcerated skin lesion. In this context, determining the region of the lesion with the highest parasite density and, consequently, the appropriate site for collecting samples can define the success of the laboratory diagnosis. Hence, this study aims to comparatively evaluate the parasite load by qPCR, quantification of amastigotes forms in the direct exam, and the histopathological profile on the inner and outer edges of ulcerated CL lesions.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec002\">\n<jats:title>Methods<\/jats:title>\n<jats:p>Samples from ulcerated skin lesions from 39 patients with confirmed CL were examined. We performed scraping of the ulcer inner edge (base) and outer edge (raised border) and lesion biopsy for imprint and histopathological examination. Slides smears were stained by Giemsa and observed in optical microscopy, the material contained on the smears was used to determine parasite load by quantitative real-time PCR (qPCR) with primers directed to the <jats:italic>Leishmania (Viannia)<\/jats:italic> minicircle kinetoplast DNA. The histopathological exam was performed to evaluate cell profile, tissue alterations and semi-quantitative assessment of amastigote forms in inner and outer edges.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec003\">\n<jats:title>Principal findings<\/jats:title>\n<jats:p>Parasite loads were higher on the inner edge compared to the outer edge of the lesions, either by qPCR technique (<jats:italic>P<\/jats:italic>&lt;0.001) and histopathological examination (<jats:italic>P<\/jats:italic>&lt; 0.003). There was no significant difference in the parasite load between the imprint and scraping on the outer edge (<jats:italic>P<\/jats:italic> = 1.0000).<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec004\">\n<jats:title>Conclusion\/Significance<\/jats:title>\n<jats:p>The results suggest that clinical specimens from the inner edge of the ulcerated CL lesions are the most suitable for both molecular diagnosis and direct parasitological examination.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pone.0243978","type":"journal-article","created":{"date-parts":[[2021,1,21]],"date-time":"2021-01-21T18:22:02Z","timestamp":1611253322000},"page":"e0243978","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":18,"title":["Comparison of parasite load by qPCR and histopathological changes of inner and outer edge of ulcerated cutaneous lesions of cutaneous leishmaniasis"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8120-1864","authenticated-orcid":true,"given":"Caio","family":"Thomaz","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5606-9854","authenticated-orcid":true,"given":"Cintia Xavier","family":"de Mello","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ot\u00e1vio de Melo","family":"Esp\u00edndola","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3460-1210","authenticated-orcid":true,"given":"Armando de Oliveira","family":"Shubach","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leonardo Pereira","family":"Quintella","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raquel Vasconcelos Carvalhaes","family":"de Oliveira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8764-7912","authenticated-orcid":true,"given":"Adriane Corr\u00eaa Gomes","family":"Duarte","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria In\u00eas Fernandes","family":"Pimentel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcelo Rosandiski","family":"Lyra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mauro Celio de Almeida","family":"Marzochi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,1,21]]},"reference":[{"key":"pone.0243978.ref001","doi-asserted-by":"crossref","first-page":"750","DOI":"10.12688\/f1000research.11120.1","article-title":"Leishmaniasis: a review","volume":"6","author":"E Torres-Guerrero","year":"2017","journal-title":"F1000Research"},{"key":"pone.0243978.ref002","volume-title":"Control of the leishmaniases: report of a meeting of the WHO Expert Commitee on the Control of Leishmaniases","author":"WHO Expert Committee on the Control of the Leishmaniases & World Health Organization","year":"2010"},{"key":"pone.0243978.ref003","volume-title":"Manual de Vigil\u00e2ncia da Leishmaniose Tegumentar Americana","author":"Ministry of Health of Brazil","year":"2017"},{"issue":"4","key":"pone.0243978.ref004","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1111\/j.1365-2133.2006.07365.x","article-title":"Unusual histopathological features of cutaneous leishmaniasis identified by polymerase chain reaction specific for Leishmania on paraffin-embedded skin biopsies","volume":"155","author":"A Boer","year":"2006","journal-title":"British Journal of Dermatology"},{"issue":"6","key":"pone.0243978.ref005","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1590\/S0036-46651986000600008","article-title":"Histopatologia da leishmaniose tegumentar por Leishmania braziliensis braziliensis: 4. 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