{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T03:52:43Z","timestamp":1773287563022,"version":"3.50.1"},"reference-count":62,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,10,28]],"date-time":"2019-10-28T00:00:00Z","timestamp":1572220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2019,10,28]],"date-time":"2019-10-28T00:00:00Z","timestamp":1572220800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Infect Dis"],"published-print":{"date-parts":[[2019,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n              <jats:sec>\n                <jats:title>Background<\/jats:title>\n                <jats:p><jats:italic>Leishmania infantum<\/jats:italic>, the etiological agent of visceral leishmaniasis, is a neglected zoonosis that requires validation and standardization of satisfactory diagnostic methodologies. Thus, the aim of the present study was to evaluate the effectiveness of cathepsin L-<jats:italic>like<\/jats:italic> protease as a target for making molecular diagnoses and as a phylogenetic marker enabling to understand the intraspecies variations and evolutionary history of <jats:italic>L. infantum<\/jats:italic> in Brazil.<\/jats:p>\n              <\/jats:sec>\n              <jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>We used 44 isolates of <jats:italic>L. infantum<\/jats:italic>. The cathepsin L-<jats:italic>like<\/jats:italic> gene fragments were amplified, sequenced, manually aligned and analyzed using inference methods. The sequences generated were used to search and design oligonucleotide primers to be used in reactions specific to the target parasite.<\/jats:p>\n              <\/jats:sec>\n              <jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>The cathepsin L-<jats:italic>like<\/jats:italic> gene did not show any intraspecies variability among the isolates analyzed. The pair of primers proposed amplified the target deoxyribonucleic acid (DNA) of <jats:italic>L. infantum<\/jats:italic> isolates and were effective for DNA amplification at concentrations of as low as 10<jats:sup>\u2212\u200911<\/jats:sup>\u2009ng\/\u03bcl. The proposed marker did not present cross-reactions with other hemoparasites. When used for making the diagnosis in a panel of clinical samples from dogs, a positivity rate of 49.03% (102\/208) was obtained, versus 14.42% (30\/208) for a ribosomal internal transcribed spacer (ITS) marker. In samples from sandflies, the rate was 6.25% and from humans, 14.28%.<\/jats:p>\n              <\/jats:sec>\n              <jats:sec>\n                <jats:title>Conclusion<jats:italic>s<\/jats:italic>\n                        <\/jats:title>\n                <jats:p>The results described in this work allow us to infer that CatLeish-PCR is a sensitive and specific marker for use in diagnostic trials of <jats:italic>L. infantum<\/jats:italic> and in clinical and epidemiological surveys.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s12879-019-4463-8","type":"journal-article","created":{"date-parts":[[2019,10,28]],"date-time":"2019-10-28T13:10:58Z","timestamp":1572268258000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Exploring Leishmania infantum cathepsin as a new molecular marker for phylogenetic relationships and visceral leishmaniasis diagnosis"],"prefix":"10.1186","volume":"19","author":[{"given":"Ryan Emiliano","family":"da Silva","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bruna Matarucco","family":"Sampaio","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Renata","family":"Tonhosolo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrea Perei ra","family":"da Costa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luiz Eduardo","family":"da Silva Costa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fernanda Ap.","family":"Nieri-Bastos","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M\u00e1rcia Aparecida","family":"Speran\u00e7a","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0478-6771","authenticated-orcid":false,"given":"Arlei","family":"Marcili","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2019,10,28]]},"reference":[{"key":"4463_CR1","first-page":"68","volume-title":"Progress in Protozoology","author":"BM Honigberg","year":"1963","unstructured":"Honigberg BM. A contribution to systematics of the non-pigmented flagellates. In: Ludvik J, Lom J, Vavra J, editors. Progress in Protozoology. New York: Academic Press; 1963. p. 68\u20139."},{"key":"4463_CR2","first-page":"213","volume-title":"Biology of the Kinetoplastida","author":"FG Wallace","year":"1979","unstructured":"Wallace FG. Biology of the Kinetoplastida of Arthropods. In: Lumsden WHR, Evans DA, editors. Biology of the Kinetoplastida. New York: Academic Press; 1979. p. 213\u201340."},{"key":"4463_CR3","doi-asserted-by":"publisher","first-page":"1317","DOI":"10.1016\/0020-7519(94)90198-8","volume":"24","author":"K Vickerman","year":"1994","unstructured":"Vickerman K. The evolutionary expansion of the trypanosomatid flagellates. Int J Parasitol. 1994;24:1317\u201331.","journal-title":"Int J Parasitol"},{"key":"4463_CR4","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1051\/parasite\/2017002","volume":"24","author":"F Galvis-Ovallos","year":"2017","unstructured":"Galvis-Ovallos F, Silva MD, Bispo GB, Oliveira AG, Neto JR, Malafronte RD, et al. Canine visceral leishmaniasis in the metropolitan area of S\u00e3o Paulo: Pintomyia fischeri as potential vector of Leishmania infantum. Parasite. 2017;24:2.","journal-title":"Parasite"},{"key":"4463_CR5","first-page":"419","volume":"45","author":"LM Deane","year":"1954","unstructured":"Deane LM, Deane MP. Encontro de Leishmanias nas v\u00edsceras e na pele de uma raposa, em zona end\u00eamica de calazar, nos arredores de Sobral, Cear\u00e1. Hospital. 1954;45:419\u201321.","journal-title":"Hospital"},{"key":"4463_CR6","doi-asserted-by":"publisher","first-page":"656","DOI":"10.1089\/vbz.2015.1771","volume":"15","author":"AP Costa","year":"2015","unstructured":"Costa AP, Costa FB, Soares HS, Ramirez DG, Mesquita ET, Gennari SM, et al. Trypanosoma cruzi and Leishmania infantum chagasi infection in wild mammals from Maranh\u00e3o state, Brazil. Vector Borne Zoonotic Dis. 2015;15:656\u201366.","journal-title":"Vector Borne Zoonotic Dis"},{"key":"4463_CR7","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1016\/j.biochi.2016.11.015","volume":"133","author":"CB Gomes","year":"2016","unstructured":"Gomes CB, Silva FS, Charret KS, Pereira BAS, Finkelstein LC, Souza RS, et al. Increasing in cysteine proteinase B expression and enzymatic activity during in vitro differentiation of Leishmania (Viannia) braziliensis: first evidence of modulation during morphological transition. Biochimie. 2016;133:28\u201336.","journal-title":"Biochimie"},{"key":"4463_CR8","doi-asserted-by":"publisher","first-page":"1541","DOI":"10.1590\/S0100-40422010000700022","volume":"33","author":"RE Silva-L\u00f3pez","year":"2010","unstructured":"Silva-L\u00f3pez RE. Proteases de Leishmania: novos alvos para o desenvolvimento racional de f\u00e1rmacos. Qu\u00edm Nova. 2010;33:1541\u20138.","journal-title":"Qu\u00edm Nova"},{"key":"4463_CR9","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1371\/journal.pntd.0005311","volume":"11","author":"M Agallou","year":"2017","unstructured":"Agallou M, Margaroni M, Athanasiou E, Toubanaki D, Kontonikola K, Karidi K, et al. Identification of BALB\/c immune markers correlated with a partial protection to Leishmania infantum after vaccination with a rationally designed multi-epitope cysteine protease a peptide-based nanovaccine. PLoS Negl Trop Dis. 2017;11:1.","journal-title":"PLoS Negl Trop Dis"},{"key":"4463_CR10","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1186\/1471-2199-6-3","volume":"6","author":"V Mundodi","year":"2005","unstructured":"Mundodi V, Kucknoor AS, Gedamu L. Role of Leishmania (Leishmania) chagasi amastigote cysteine protease in intracelular parasite survival: studies by gene disrruption and antisense mRNA inhibition. BMC Mol Biol. 2005;6:3.","journal-title":"BMC Mol Biol"},{"key":"4463_CR11","doi-asserted-by":"publisher","first-page":"375","DOI":"10.1016\/j.mib.2004.06.010","volume":"7","author":"JC Mottram","year":"2004","unstructured":"Mottram JC, Coombs GH, Alexander J. Cysteine peptidases as virulence factors of Leishmania. Curr Opin Microbiol. 2004;7:375\u201381.","journal-title":"Curr Opin Microbiol"},{"key":"4463_CR12","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1371\/journal.ppat.1005658","volume":"12","author":"PA Casgrain","year":"2016","unstructured":"Casgrain PA, Martel C, McMaster WR, Mottram JC, Olivier M, Descoteaux A. Cysteine peptidase B regulates Leishmania mexicana virulence through the modulation of GP63 expression. PLoS Pathog. 2016;12:5.","journal-title":"PLoS Pathog"},{"key":"4463_CR13","doi-asserted-by":"publisher","first-page":"44","DOI":"10.1016\/j.mcp.2008.11.003","volume":"23","author":"AP Cortez","year":"2008","unstructured":"Cortez AP, Rodrigues AC, Garcia HA, Neves L, Batista JS, Bengaly Z, et al. Cathepsin L-like genes of Trypanosoma vivax from Africa and South America characterization, relationships and diagnostic implications. Mol Cell Probes. 2008;23:44\u201351.","journal-title":"Mol Cell Probes"},{"key":"4463_CR14","doi-asserted-by":"publisher","first-page":"363","DOI":"10.1016\/j.vetpar.2011.03.017","volume":"180","author":"HA Garcia","year":"2011","unstructured":"Garcia HA, Kamyingkird K, Rodrigues AC, Jittapalapong S, Teixeira MM, Desquesnes M. High genetic diversity in field isolates of Trypanosoma theileri assessed by analysis of cathepsin L-like sequences disclosed multiple and new genotypes infecting cattle in Thailand. Vet Parasitol. 2011;180:363\u20137.","journal-title":"Vet Parasitol"},{"key":"4463_CR15","doi-asserted-by":"publisher","first-page":"335","DOI":"10.1016\/j.vetpar.2014.12.006","volume":"207","author":"N Yokoyama","year":"2015","unstructured":"Yokoyama N, Sivakumar T, Fukushi S, Tattiyapong M, Tuvshintulga B, Kothalawala H, et al. Genetic diversity in Trypanosoma theileri from Sri Lankan cattle and water buffaloes. Vet Parasitol. 2015;207:335\u201341.","journal-title":"Vet Parasitol"},{"key":"4463_CR16","unstructured":"Ferrer L. Clinical aspects of canine leishmaniasis. In: R Killick-Kendrick, editor. Canine leishmaniasis: an updat. Proceedings of a Canine Leishmaniasis Forum. Barcelona: Hoeschst Roussel Vet; 1999. p. 6\u201310."},{"key":"4463_CR17","first-page":"107","volume":"17","author":"TSM Assis","year":"2008","unstructured":"Assis TSM, Braga ASC, Pedras MJ, Barral AMP, Siqueira IC, Costa CHN, et al. Valida\u00e7\u00e3o do teste imunocromatogr\u00e1fico r\u00e1pido IT-LEISH\u00ae para o diagn\u00f3stico da leishmaniose visceral humana. Epidemiol Serv Sa\u00fade Bras\u00edlia. 2008;17:107\u201316.","journal-title":"Epidemiol Serv Sa\u00fade Bras\u00edlia"},{"key":"4463_CR18","first-page":"13","volume":"6","author":"MD Laurenti","year":"2009","unstructured":"Laurenti MD. Correla\u00e7\u00e3o entre o diagn\u00f3stico parasitol\u00f3gico e sorol\u00f3gico na leishmaniose visceral americana canina. Bol Epidemiol Paul. 2009;6:13\u201323.","journal-title":"Bol Epidemiol Paul"},{"key":"4463_CR19","doi-asserted-by":"publisher","first-page":"775","DOI":"10.1073\/pnas.90.2.775","volume":"90","author":"JM Burns","year":"1993","unstructured":"Burns JM, Shreffler WG, Benson DR, Ghalib HW, Badaro R, Reed SG. Molecular characterization of a kinesin-related antigen of Leishmania chagasi that detects specific antibody in African and American visceral leishmaniasis. Proc Natl Acad Sci. 1993;90:775\u20139.","journal-title":"Proc Natl Acad Sci"},{"key":"4463_CR20","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/0304-4017(96)00946-6","volume":"65","author":"F Mancianti","year":"1996","unstructured":"Mancianti F, Pedonese F, Poli A. Evaluation of dot enzyme-linked immunosorbent assay (dot-ELISA) for the serodiagnosis of canine leishmaniosis as compared with indirect immunofluorescence assay. Vet Parasitol. 1996;65:1\u20139.","journal-title":"Vet Parasitol"},{"key":"4463_CR21","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1016\/j.vetpar.2006.09.017","volume":"144","author":"E Ferroglio","year":"2006","unstructured":"Ferroglio E, Centaro E, Mignone W, Trisciuoglio A. Evaluation of an ELISA rapid device for the serological diagnosis of Leishmania infantum infection in dog as compared with immunofluorescence assay and Western blot. Vet Parasitol. 2006;144:162\u20136.","journal-title":"Vet Parasitol"},{"key":"4463_CR22","doi-asserted-by":"publisher","first-page":"33","DOI":"10.5216\/rpt.v44i1.34799","volume":"44","author":"LC Hirschmann","year":"2015","unstructured":"Hirschmann LC, Brod CS, Radin J, Simon CF, Recuero AL. Leishmaniose visceral canina: compara\u00e7\u00e3o de m\u00e9todos sorol\u00f3gicos em c\u00e3es de area indene do Rio Grande do Sul no Brasil. Rev Patol Trop. 2015;44:33\u201344.","journal-title":"Rev Patol Trop"},{"key":"4463_CR23","doi-asserted-by":"publisher","first-page":"32","DOI":"10.1186\/1756-3305-6-32","volume":"6","author":"BB Yanh","year":"2013","unstructured":"Yanh BB, Chen DL, Chen JP, Liao L, Hu XS, Xu JN. Analysis of kinetoplast cytochrome b gene of 16 Leishmania isolates from different foci of China: different species of Leishmania in China and their phylogenetic inference. Parasit Vectors. 2013;6:32.","journal-title":"Parasit Vectors"},{"key":"4463_CR24","doi-asserted-by":"publisher","first-page":"283","DOI":"10.1016\/j.exppara.2006.09.011","volume":"115","author":"T Kuru","year":"2007","unstructured":"Kuru T, Jirata D, Genetu A, Barr S, Mengistu Y, Aseffa A, et al. Leishmania aethiopica: identification and characterization of cathepsin L-like cysteine protease genes. Exp Parasitol. 2007;115:283\u201390.","journal-title":"Exp Parasitol"},{"key":"4463_CR25","doi-asserted-by":"publisher","first-page":"4876","DOI":"10.1093\/nar\/25.24.4876","volume":"25","author":"JD Thompson","year":"1997","unstructured":"Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. The clustalX windons interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 1997;25:4876\u201382.","journal-title":"Nucleic Acids Res"},{"key":"4463_CR26","first-page":"24","volume":"4","author":"KB Nicholas","year":"1997","unstructured":"Nicholas KB, Nicholas HB, Deerfield DW, Nicholas HBJ, DWI DWID, Nicholas KB, et al. GeneDoc: analysis and visualization of genetic variation. Embnewnews. 1997;4:24.","journal-title":"Embnewnews"},{"key":"4463_CR27","volume-title":"PAUP: phylogenetic analysis using parsimony. Version 4b6","author":"DL Swofford","year":"1998","unstructured":"Swofford DL. PAUP: phylogenetic analysis using parsimony. Version 4b6. Suderland: Sinauer Associates; 1998."},{"key":"4463_CR28","doi-asserted-by":"publisher","first-page":"754","DOI":"10.1093\/bioinformatics\/17.8.754","volume":"17","author":"JP Huelsenbeck","year":"2001","unstructured":"Huelsenbeck JP, Ronquist F. MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics. 2001;17:754\u20135.","journal-title":"Bioinformatics"},{"key":"4463_CR29","doi-asserted-by":"publisher","first-page":"575","DOI":"10.1016\/S0035-9203(00)90093-2","volume":"94","author":"NO El Tai","year":"2000","unstructured":"El Tai NO, Osman OF, El Fari M, Presber W, Sch\u00f6nian G. Genetic heterogeneity of ribosomal internal transcribed spacer in clinical samples of Leishmania donovani spotted on filter paper as revealed by single-strand conformation polymorphisms and sequencing. Trans R Soc Trop Med Hyg. 2000;94:575\u20139.","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"4463_CR30","doi-asserted-by":"publisher","first-page":"44","DOI":"10.1016\/j.meegid.2014.04.001","volume":"25","author":"A Marcili","year":"2014","unstructured":"Marcili A, Speran\u00e7a MA, Costa AP, Madeira MF, Soares HS, Sanches CO, et al. Phylogenetic relationships of Leishmania species based on trypanosomatid barcode (SSU rDNA) and gGAPDH genes: taxonomic revision of Leishmania (L.) infantum chagasi in South America. Infect Genet Evol. 2014;25:44\u201351.","journal-title":"Infect Genet Evol"},{"key":"4463_CR31","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1371\/journal.pntd.0004349","volume":"10","author":"M Akhoundi","year":"2016","unstructured":"Akhoundi M, Kuhls K, Cannet A, Vot\u00fdpka J, Marty P, Delaunay P, et al. A historical overview of the classification, evolution, and dispersion of Leishmania parasites and sandflies. PLoS Negl Trop Dis. 2016;10:3.","journal-title":"PLoS Negl Trop Dis"},{"key":"4463_CR32","doi-asserted-by":"publisher","first-page":"188","DOI":"10.1016\/S0169-4758(00)01637-9","volume":"16","author":"IL Maur\u00edcio","year":"2000","unstructured":"Maur\u00edcio IL, Stothard JR, Miles MA. The strange case of Leishmania chagasi. Parasitol Today. 2000;16:188\u20139.","journal-title":"Parasitol Today"},{"key":"4463_CR33","first-page":"1","volume-title":"The Leishmaniases in biology and medicine","author":"R Lainson","year":"1978","unstructured":"Lainson R, Shaw JJ. Evolution, classification and geographic distribution. In: Peters W, Killick-Kendrick R, editors. The Leishmaniases in biology and medicine. London: Academic Press; 1978. p. 1\u2013120."},{"key":"4463_CR34","doi-asserted-by":"publisher","first-page":"275","DOI":"10.1016\/S0035-9203(98)91009-4","volume":"92","author":"BL Travi","year":"1998","unstructured":"Travi BL, Osorio Y, Becerra MT, Adler GH. Dynamics of Leishmania chagasi infection in small mammals of the undisturbed and degraded tropical dry forests of northern Colombia. Trans R Soc Trop Med Hyg. 1998;92:275\u20138.","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"4463_CR35","doi-asserted-by":"publisher","first-page":"651","DOI":"10.1080\/00034983.2000.11813588","volume":"94","author":"AM D\u00e1vila","year":"2000","unstructured":"D\u00e1vila AM, Momen H. Internal-transcribed-spacer (ITS) sequences used to explore phylogenetic relationships within Leishmania. Ann Trop Med Parasitol. 2000;94:651\u20134.","journal-title":"Ann Trop Med Parasitol"},{"key":"4463_CR36","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1017\/S0031182004004792","volume":"128","author":"GE Luyo-Acero","year":"2004","unstructured":"Luyo-Acero GE, Uezato H, Oshiro M, Takei K, Kariya K, Katakura K, et al. Sequence variation of the cytochrome b gene of various human infecting members of the genus Leishmania and their phylogeny. Parasitology. 2004;128:483\u201391.","journal-title":"Parasitology"},{"key":"4463_CR37","first-page":"362","volume":"12","author":"MA Fornazieri","year":"2008","unstructured":"Fornazieri MA, Yamaguti HY, Moreira JH, Takemoto LE, Navarro PL, Hesbike RE. The Most common Otorhinolaryngologic manifestations of granulomatous diseases. Int Arch Otorhinolaryngol. 2008;12:362\u20135.","journal-title":"Int Arch Otorhinolaryngol"},{"key":"4463_CR38","doi-asserted-by":"publisher","first-page":"214","DOI":"10.1111\/j.1468-3083.1998.tb00738.x","volume":"10","author":"IS Souza","year":"1998","unstructured":"Souza IS, Naiff RD, Guimar\u00e3es TC, Naiff MF, Cupolilo E, Rodrigues WA, et al. American cutaneous leishmaniasis due to Leishmania (Viannia) guyanensis as an initial clinical presentation of human immunodeficiency virus infection. J Eur Acad Dermatol Venereol. 1998;10:214\u20137.","journal-title":"J Eur Acad Dermatol Venereol"},{"key":"4463_CR39","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1080\/00034983.1912.11687069","volume":"6","author":"H Seidelin","year":"1912","unstructured":"Seidelin H. Leishmaniasis and Babesiasis in Yucatan. Ann Trop Med Parasitol. 1912;6:295\u2013300.","journal-title":"Ann Trop Med Parasitol"},{"key":"4463_CR40","doi-asserted-by":"publisher","first-page":"735","DOI":"10.1016\/0035-9203(91)90437-4","volume":"85","author":"FT Silveira","year":"1991","unstructured":"Silveira FT, Lainson R, Shaw JJ, Souza AA, Ishikawa EA, Braga RR. Cutaneous leishmaniasis due to Leishmania (Leishmania) amazonensis in Amazonian Brazil, and the significance of a negative Montenegro skin-test in human infections. Trans R Soc Trop Med Hyg. 1991;85:735\u20138.","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"4463_CR41","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1016\/S0001-706X(98)00049-7","volume":"71","author":"E Martinez","year":"1998","unstructured":"Martinez E, Le Pont F, Torrez M, Telleria J, Vargas F, Mu\u00f1oz M, et al. A new focus of cutaneous leishmaniasis due to Leishmania amazonensis in a sub Andean region of Bolivia. Acta Trop. 1998;71:97\u2013106.","journal-title":"Acta Trop"},{"key":"4463_CR42","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1590\/S0036-46652005000400003","volume":"47","author":"FJA Narvaez","year":"2005","unstructured":"Narvaez FJA, Medina-Peralta S, Vargas-Gonzalez A, Canto-Lara SB, Estrada-Parra S. The histopathology of cutaneous leishmaniasis due to Leishmania (Leishmania) Mexicana in the Yucatan peninsula, Mexico. Rev Inst Med Trop Sao Paulo. 2005;47:191\u20134.","journal-title":"Rev Inst Med Trop Sao Paulo"},{"key":"4463_CR43","first-page":"147","volume":"8","author":"JAL Lindoso","year":"2016","unstructured":"Lindoso JAL, Cunha MA, Queiroz IT, Moreira CHV. Leishmaniasis\u2013HIV coinfection: current challenges. HIV AIDS (Auckl). 2016;8:147\u201356.","journal-title":"HIV AIDS (Auckl)"},{"key":"4463_CR44","doi-asserted-by":"publisher","first-page":"6597","DOI":"10.1128\/IAI.70.12.6597-6605.2002","volume":"70","author":"M Colmenares","year":"2002","unstructured":"Colmenares M, Constant SL, Kima PE, McMahon-Pratt D. Leishmania pifanoi pathogenesis: selective lack of a local cutaneous response in the absence of circulating antibody. Infect Immun. 2002;70:6597\u2013605.","journal-title":"Infect Immun"},{"key":"4463_CR45","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1155\/2012\/279081","volume":"2012","author":"Omar Hern\u00e1ndez-Montes","year":"2012","unstructured":"Montes OH, Guzm\u00e1n SG, G\u00f3mez FM, Barker DC, Monroy-Ostria A. Analysis of Kinetoplast DNA from Mexican Isolates of Leishmania (L.) mexicana. Interdiscip Perspect Infect Dis. 2012. \n                    https:\/\/doi.org\/10.1155\/2012\/279081\n                    \n                  .","journal-title":"Interdisciplinary Perspectives on Infectious Diseases"},{"key":"4463_CR46","doi-asserted-by":"publisher","first-page":"684","DOI":"10.1128\/EC.00405-07","volume":"7","author":"C Mendonza-Palomares","year":"2008","unstructured":"Mendonza-Palomares C, Biteau N, Giroud C, Coustou V, Coetzer T, Authi\u00e9 E, et al. Molecular and biochemical characterization of a Cathepsin B-like protease family unique to Trypanosoma congolense. Eukaryot Cell. 2008;7:684\u201397.","journal-title":"Eukaryot Cell"},{"key":"4463_CR47","doi-asserted-by":"publisher","first-page":"318","DOI":"10.1016\/j.parint.2010.03.002","volume":"58","author":"AC Rodrigues","year":"2010","unstructured":"Rodrigues AC, Garcia HA, Ortiz PA, Cortez AP, Martinkovic F, Paiva F, et al. Cysteine proteases of Trypanosoma (Megatrypanum) theileri: cathepsin L-like gene sequences as targets for phylogenetic analysis, genotyping diagnosis. Parasitol Int. 2010;58:318\u201325.","journal-title":"Parasitol Int"},{"key":"4463_CR48","first-page":"6","volume":"7","author":"L Lima","year":"2012","unstructured":"Lima L, Ortiz PA, Silva FM, Alves JM, Serrano MG, Cortez AP, et al. Repertoire, genealogy and genomic organization of cruzipain and homologous genes in Trypanosoma cruzi, T. cruzi-like and other trypanosome species. PLoS One. 2012;7:6.","journal-title":"PLoS One"},{"key":"4463_CR49","doi-asserted-by":"publisher","first-page":"249","DOI":"10.1016\/j.actatropica.2009.07.036","volume":"112","author":"PA Ortiz","year":"2009","unstructured":"Ortiz PA, Maia da Silva F, Cortez AP, Lima L, Campaner M, Pral EM, et al. Genes of cathepsin L-like proteases in Trypanosoma rangeli isolates: markers for diagnosis, genotyping and phylogenetic relationships. Acta Trop. 2009;112:249\u201359.","journal-title":"Acta Trop"},{"key":"4463_CR50","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1186\/1471-2199-7-42","volume":"7","author":"H Denise","year":"2006","unstructured":"Denise H, Poot J, Jim\u00e9nez M, Ambit A, Herrmann DC, Vermeulen AN, Coombs GH, et al. Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5. BMC Mol Biol. 2006;7:42.","journal-title":"BMC Mol Biol"},{"key":"4463_CR51","doi-asserted-by":"publisher","first-page":"160","DOI":"10.1186\/1756-3305-5-160","volume":"5","author":"M Silva-Almeida","year":"2012","unstructured":"Silva-Almeida M, Pereira BAS, Ribeiro-Guimar\u00e3es ML, Alves CR. Proteinases as virulence factors in Leishmania spp. infection in mammals. Parasit Vectors. 2012;5:160.","journal-title":"Parasit Vectors"},{"key":"4463_CR52","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1371\/journal.pntd.0004485","volume":"10","author":"RA Zampieri","year":"2016","unstructured":"Zampieri RA, Laranjeira-Silva MF, Muxel SM, Lima ACS, Shaw JJ, Floeter-Winter LM. High resolution melting analysis targeting hsp70 as a fast and efficient method for the discrimination of Leishmania species. PLoS Negl Trop Dis. 2016;10:2.","journal-title":"PLoS Negl Trop Dis"},{"key":"4463_CR53","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1016\/j.mimet.2016.10.002","volume":"131","author":"RP Silva","year":"2016","unstructured":"Silva RP, Silva LAMT, Silva MAL, Alburquerque SCG, Goes TCG, Morais RCS, Melo FL, Cavalcante MP. Evaluation of urine for Leishmania infantum DNA detection by real-time quantitative PCR. J Microbiol Methods. 2016;131:34\u201341.","journal-title":"J Microbiol Methods"},{"key":"4463_CR54","first-page":"2","volume":"1","author":"AP Costa","year":"2014","unstructured":"Costa AP, Ferreira JIGS, Fournier GFSR, Lopes MG, Ramirez D, Acosta ICL, et al. Survey of Leishmania Infantum Chagasi in wild and domestic animals in urban area and Atlantic rainforest fragment in northeast, Brazil. J Biodivers Biopros Dev. 2014;1:2.","journal-title":"J Biodivers Biopros Dev"},{"key":"4463_CR55","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1371\/journal.pntd.0004333","volume":"10","author":"DB Fraga","year":"2016","unstructured":"Fraga DB, Pacheco LV, Borja LS, Tuy PG, Bastos LA, Solc\u00e0 MS, Amorim LD, et al. The rapid test based on Leishmania infantum chimeric rK28 protein improves the diagnosis of canine visceral Leishmaniasis by reducing the detection of false-positive dogs. PLoS Negl Trop Dis. 2016;10:1.","journal-title":"PLoS Negl Trop Dis"},{"key":"4463_CR56","doi-asserted-by":"publisher","first-page":"2110","DOI":"10.1128\/JCM.02555-06","volume":"45","author":"F Foulet","year":"2007","unstructured":"Foulet F, Botterel F, Buffet P, Morizot G, Rivollet D, Deniau M, et al. Detection and identification of Leishmania species from clinical specimens by using a real-time PCR assay and sequencing of the cytochrome b gene. J Clin Microbiol. 2007;45:2110\u20135.","journal-title":"J Clin Microbiol"},{"key":"4463_CR57","first-page":"2","volume":"9","author":"M Ceccarelli","year":"2014","unstructured":"Ceccarelli M, Galluzzi L, Migliazzo A, Magnani M. Detection and characterization of Leishmania (Leishmania) and Leishmania (Viannia) by SYBR green-based real-time PCR and high resolution melt analysis targeting kinetoplast minicircle DNA. PLoS One. 2014;9:2.","journal-title":"PLoS One"},{"key":"4463_CR58","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s13071-017-2181-x","volume":"10","author":"M Ceccarelli","year":"2017","unstructured":"Ceccarelli M, Galluzzi L, Diotallevi A, Andreoni F, Fowler H, Petersen C, et al. The use of kDNA minicircle subclass relative abundance to differentiate between Leishmania (L.) infantum and Leishmania (L.) amazonensis. Parasit Vectors. 2017;10:1.","journal-title":"Parasit Vectors"},{"key":"4463_CR59","doi-asserted-by":"publisher","first-page":"65","DOI":"10.1016\/j.mcp.2016.08.006","volume":"31","author":"F Dantas-Torres","year":"2017","unstructured":"Dantas-Torres F, Sales KGS, Silva LG, Otranto D, Figueredo LA. Leishmania-FAST15: a rapid, sensitive and low-cost real-time PCR assay for the detection of Leishmania infantum and Leishmania braziliensis kinetoplast DNA in canine blood samples. Mol Cell Probes. 2017;31:65\u20139.","journal-title":"Mol Cell Probes"},{"key":"4463_CR60","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1016\/S0732-8893(03)00040-3","volume":"46","author":"J Marfurt","year":"2003","unstructured":"Marfurt J, Niederwieser I, Makia ND, Beck HP, Felger I. Diagnostic genotyping of old and New World Leishmania species by PCR-RFLP. Diagn Microbiol Infect Dis. 2003;46:115\u201324.","journal-title":"Diagn Microbiol Infect Dis"},{"key":"4463_CR61","doi-asserted-by":"publisher","first-page":"339","DOI":"10.1590\/S0365-05962005001000013","volume":"80","author":"FB Garcia","year":"2005","unstructured":"Garcia FB, Santos SSR, Chociay MF, Medeiros ACR, Roselino AMF. M\u00e9todos subsidi\u00e1rios para o diagn\u00f3stico da Leishmaniose tegumentar americana (LTA): compara\u00e7\u00e3o dos resultados do seq\u00fcenciamento de DNA e da PCR-RFLP para determina\u00e7\u00e3o da esp\u00e9cie de Leishmania em amostras cut\u00e2neo-mucosas. An Bras. 2005;80:339\u201344.","journal-title":"An Bras"},{"key":"4463_CR62","first-page":"215","volume":"12","author":"MA Mohammadi","year":"2017","unstructured":"Mohammadi MA, Bamorovat M, Harandi MF, Karimi T, Sharifi I, Aflatoonian MR. Comparison of three PCR-based methods for simplicity and cost effectiveness identification of cutaneous Leishmaniasis due to Leishmania tropica. Iran J Parasitol. 2017;12:215\u201323.","journal-title":"Iran J Parasitol"}],"container-title":["BMC Infectious Diseases"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/link.springer.com\/content\/pdf\/10.1186\/s12879-019-4463-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/link.springer.com\/article\/10.1186\/s12879-019-4463-8\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/link.springer.com\/content\/pdf\/10.1186\/s12879-019-4463-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,10,27]],"date-time":"2020-10-27T00:12:26Z","timestamp":1603757546000},"score":1,"resource":{"primary":{"URL":"https:\/\/bmcinfectdis.biomedcentral.com\/articles\/10.1186\/s12879-019-4463-8"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,10,28]]},"references-count":62,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2019,12]]}},"alternative-id":["4463"],"URL":"https:\/\/doi.org\/10.1186\/s12879-019-4463-8","relation":{},"ISSN":["1471-2334"],"issn-type":[{"value":"1471-2334","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,10,28]]},"assertion":[{"value":"21 February 2019","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"11 September 2019","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"28 October 2019","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The human samples were transferred by the Laboratory of Pathogens from Center of Natural and Human Sciences from Universidade Federal do ABC and approved by Ethics Committee for Human research from Federal University of ABC (number 883722718900005594). The clinical samples from dogs and sandflies procedures were endorsed by the Ethics Committee for Animal Use of the University of S\u00e3o Paulo, Brazil (number 7584250216) according National Council for Control of Animal Experimentation.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare that they have no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"895"}}