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In this context, serological evaluation is essential for Canine leishmaniasis management. Several serological alternatives, such as rapid diagnostic tests, enzyme-linked immunosorbent assay (ELISA) and immunofluorescence antibody test (IFAT), are well established. In fact, the capacity of distinct tests and antigens, evaluated by their sensitivity and specificity, to detect disease is normally considered sufficient for diagnosing Canine leishmaniasis. In this context, we evaluated the seropositivity using 8 different serological tests (ELISA with <jats:italic>Leishmania<\/jats:italic> recombinant proteins (rK39, LicTXNPx); soluble promastigote <jats:italic>Leishmania<\/jats:italic> antigens (SPLA); commercial ELISA test) in 82 clinically suspect animals from Northern Portugal. The obtained serological data originated 50% of inconclusive serological information with a mixture of seropositive and seronegative results for individual animals. Cut-off independent risk groups were then generated from the serological data to evaluate the clustering of the samples. This analysis originated risk groups that correlated with the most seropositive samples, suggesting that this method might be used, in a cut-off independent manner, to improve conventional serological evaluation. Ultimately, given that no test prioritization exists, the use of any single serological test increases the potential for misdiagnosis, along with all associated risks for the dog as well as public health. The use of a cut-off independent analysis has the potential to improve the predictive values of these tests, enabling a more accurate evaluation of the dog\u2019s condition.<\/jats:p>","DOI":"10.1038\/s41598-020-60067-6","type":"journal-article","created":{"date-parts":[[2020,2,20]],"date-time":"2020-02-20T11:03:11Z","timestamp":1582196591000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Challenges in the serological evaluation of dogs clinically suspect for canine leishmaniasis"],"prefix":"10.1038","volume":"10","author":[{"given":"Nuno","family":"Santar\u00e9m","sequence":"first","affiliation":[]},{"given":"Susana","family":"Sousa","sequence":"additional","affiliation":[]},{"given":"C\u00e9lia G.","family":"Amorim","sequence":"additional","affiliation":[]},{"given":"Nuno Lima","family":"de Carvalho","sequence":"additional","affiliation":[]},{"given":"Hugo Lima","family":"de Carvalho","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9010-2075","authenticated-orcid":false,"given":"\u00d3scar","family":"Felgueiras","sequence":"additional","affiliation":[]},{"given":"Margarida","family":"Brito","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2392-6087","authenticated-orcid":false,"given":"Anabela Cordeiro","family":"da Silva","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2020,2,20]]},"reference":[{"key":"60067_CR1","doi-asserted-by":"publisher","first-page":"425","DOI":"10.1016\/j.tvjl.2014.09.002","volume":"202","author":"C Noli","year":"2014","unstructured":"Noli, C. & Saridomichelakis, M. N. An update on the diagnosis and treatment of canine leishmaniosis caused by Leishmania infantum (syn. L. chagasi). Vet. J. 202, 425\u2013435, https:\/\/doi.org\/10.1016\/j.tvjl.2014.09.002 (2014).","journal-title":"Vet. J."},{"key":"60067_CR2","doi-asserted-by":"publisher","first-page":"552","DOI":"10.1111\/vcp.12413","volume":"45","author":"S Paltrinieri","year":"2016","unstructured":"Paltrinieri, S., Gradoni, L., Roura, X., Zatelli, A. & Zini, E. Laboratory tests for diagnosing and monitoring canine leishmaniasis. Vet. Clin. Pathol. 45, 552\u2013578, https:\/\/doi.org\/10.1111\/vcp.12413 (2016).","journal-title":"Vet. Clin. Pathol."},{"key":"60067_CR3","doi-asserted-by":"publisher","first-page":"1184","DOI":"10.2460\/javma.236.11.1184","volume":"236","author":"S Paltrinieri","year":"2010","unstructured":"Paltrinieri, S. et al. Guidelines for diagnosis and clinical classification of leishmaniasis in dogs. J. Am. Vet. Med. 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