{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T05:21:32Z","timestamp":1785216092902,"version":"3.55.0"},"reference-count":83,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2024,4,25]],"date-time":"2024-04-25T00:00:00Z","timestamp":1714003200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"name":"Global Emerging Infections Surveillance","award":["ProMIS ID 17_N6_1.1.3, 2017-2018"],"award-info":[{"award-number":["ProMIS ID 17_N6_1.1.3, 2017-2018"]}]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:sec id=\"sec001\">\n<jats:title>Background<\/jats:title>\n<jats:p>Accuracy of molecular tools for the identification of parasites that cause human cutaneous leishmaniasis (CL) could largely depend on the sampling method. Non-invasive or less-invasive sampling methods such as filter paper imprints and cotton swabs are preferred over punch biopsies and lancet scrapings for detection methods of <jats:italic>Leishmania<\/jats:italic> based on polymerase chain reaction (PCR) because they are painless, simple, and inexpensive, and of benefit to military and civilian patients to ensure timely treatment. However, different types of samples can generate false negatives and there is a clear need to demonstrate which sample is more proper for molecular assays.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec002\">\n<jats:title>Methodology<\/jats:title>\n<jats:p>Here, we compared the sensitivity of molecular identification of different <jats:italic>Leishmania (Viannia)<\/jats:italic> species from Peru, using three types of sampling: punch biopsy, filter paper imprint and lancet scraping. Different composite reference standards and latent class models allowed to evaluate the accuracy of the molecular tools. Additionally, a quantitative PCR assessed variations in the results and parasite load in each type of sample.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec003\">\n<jats:title>Principal findings<\/jats:title>\n<jats:p>Different composite reference standards and latent class models determined higher sensitivity when lancet scrapings were used for sampling in the identification and determination of <jats:italic>Leishmania (Viannia)<\/jats:italic> species through PCR-based assays. This was consistent for genus identification through kinetoplastid DNA-PCR and for the determination of species using FRET probes-based Nested Real-Time PCR. Lack of species identification in some samples correlated with the low intensity of the PCR electrophoretic band, which reflects the low parasite load in samples.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec004\">\n<jats:title>Conclusions<\/jats:title>\n<jats:p>The type of clinical sample can directly influence the detection and identification of <jats:italic>Leishmania (Viannia)<\/jats:italic> species. Here, we demonstrated that lancet scraping samples consistently allowed the identification of more leishmaniasis cases compared to filter paper imprints or biopsies. This procedure is inexpensive, painless, and easy to implement at the point of care and avoids the need for anesthesia, surgery, and hospitalization and therefore could be used in resource limited settings for both military and civilian populations.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pntd.0012113","type":"journal-article","created":{"date-parts":[[2024,4,25]],"date-time":"2024-04-25T17:20:46Z","timestamp":1714065646000},"page":"e0012113","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":5,"title":["Sampling is decisive to determination of Leishmania (Viannia) species"],"prefix":"10.1371","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3379-4869","authenticated-orcid":true,"given":"Maxy B.","family":"De los Santos","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steev","family":"Loyola","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erika S.","family":"Perez-Velez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rocio del Pilar","family":"Santos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ivonne Melissa","family":"Ram\u00edrez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hugo O.","family":"Valdivia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,4,25]]},"reference":[{"key":"pntd.0012113.ref001","doi-asserted-by":"crossref","first-page":"1224","DOI":"10.1111\/j.1468-3083.2011.04266.x","article-title":"Chronic cutaneous leishmaniasis, a great mimicker with various clinical presentations: 12 years experience from Aleppo.","volume":"26","author":"MD Douba","year":"2012","journal-title":"J Eur Acad Dermatology Venereol"},{"key":"pntd.0012113.ref002","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1016\/S1473-3099(07)70209-8","article-title":"Cutaneous leishmaniasis.","author":"R Reithinger","year":"2007","journal-title":"Lancet Infectious Diseases"},{"key":"pntd.0012113.ref003","first-page":"99","article-title":"Cutaneous Leishmaniasis: Recent Developments in Diagnosis and Management.","volume-title":"American Journal of Clinical Dermatology","author":"HJC de Vries","year":"2015"},{"key":"pntd.0012113.ref004","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.1086\/648730","article-title":"Detection and Species Identification of Leishmania DNA from Filter Paper Lesion Impressions for Patients with American Cutaneous Leishmaniasis","volume":"50","author":"AK Boggild","year":"2010","journal-title":"Clin Infect Dis"},{"key":"pntd.0012113.ref005","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":"E Bensoussan","year":"2006","journal-title":"J Clin Microbiol"},{"key":"pntd.0012113.ref006","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":"WR Faber","year":"2003","journal-title":"J Am Acad Dermatol"},{"key":"pntd.0012113.ref007","doi-asserted-by":"crossref","first-page":"895","DOI":"10.4269\/ajtmh.13-0582","article-title":"Comparison of clinical samples and methods in chronic cutaneous leishmaniasis","volume":"91","author":"F Eroglu","year":"2014","journal-title":"Am J Trop Med Hyg"},{"key":"pntd.0012113.ref008","unstructured":"WHO. 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