{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T16:23:09Z","timestamp":1792513389427,"version":"4.5.0"},"reference-count":26,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2021,3,16]],"date-time":"2021-03-16T00:00:00Z","timestamp":1615852800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Federal Ministry of Education and Research, Germany, and the Ministry of Foreign Affairs, Government of the Netherlands, through FIND. RD16CIII\/0003\/0002 from the Red de Investigaci\u00f3n Cooperativa de Enfermedades Tropicales, Subprograma RETICS del Plan Est","award":["RD16CIII\/0003\/0002"],"award-info":[{"award-number":["RD16CIII\/0003\/0002"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Loop-mediated isothermal amplification allows the rapid, sensitive and specific amplification of DNA without complex and expensive equipment. We compared the diagnostic performance of Loopamp\u2122 Leishmania Detection Kit (Eiken Chemical Co., Ltd., Tokyo, Japan) with conventional and real-time polymerase chain reaction (PCR) for human cutaneous and visceral leishmaniasis caused by L. infantum. A total of 230 DNA samples from cutaneous (CL) and visceral (VL) leishmaniasis cases and controls from Spain, characterized by Leishmania nested PCR (LnPCR) were tested by: (i) the Loopamp\u2122 Leishmania Detection Kit (Loopamp), run on Genie III real-time fluorimeter (OptiGene, UK); and (ii) real-time quantitative PCR (qPCR). The Loopamp test returned 98.8% (95% confidence interval\u2014CI: 96.0\u2013100.00) sensitivity and specificity of 97.7% (95% CI: 92.2\u2013100) on VL samples, and 100% (95% CI: 99.1\u2013100) sensitivity and 100.0% (95% CI: 98.8\u2013100.0) specificity on CL samples. The Loopamp time-to-positivity (Tp) obtained by real-time fluorimetry showed excellent concordance (C = 97.91%) and strong correlation (r = 0.799) with qPCR\u2019s cycle threshold (Ct). The performance of Loopamp is comparable to that of LnPCR and qPCR in the diagnosis of cutaneous and visceral leishmaniasis due to L. infantum. The excellent correlation between the Tp and Ct should be further investigated to determine the accuracy of Loopamp to quantify parasite load in tissues.<\/jats:p>","DOI":"10.3390\/microorganisms9030610","type":"journal-article","created":{"date-parts":[[2021,3,16]],"date-time":"2021-03-16T12:01:02Z","timestamp":1615896062000},"page":"610","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Loop-Mediated Isothermal Amplification Allows Rapid, Simple and Accurate Molecular Diagnosis of Human Cutaneous and Visceral Leishmaniasis Caused by Leishmania infantum When Compared to PCR"],"prefix":"10.3390","volume":"9","author":[{"given":"Ana Victoria","family":"Ibarra-Meneses","sequence":"first","affiliation":[{"name":"WHO Collaborating Centre for Leishmaniasis, National Center for Microbiology, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carmen","family":"Chicharro","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for Leishmaniasis, National Center for Microbiology, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carmen","family":"S\u00e1nchez","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for Leishmaniasis, National Center for Microbiology, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emilia","family":"Garc\u00eda","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for Leishmaniasis, National Center for Microbiology, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheila","family":"Ortega","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for Leishmaniasis, National Center for Microbiology, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph Mathu","family":"Ndung\u2019u","sequence":"additional","affiliation":[{"name":"Foundation for Innovative New Diagnostics, 1202 Geneva, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Javier","family":"Moreno","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for Leishmaniasis, National Center for Microbiology, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2200-2360","authenticated-orcid":false,"given":"Israel","family":"Cruz","sequence":"additional","affiliation":[{"name":"Foundation for Innovative New Diagnostics, 1202 Geneva, Switzerland"},{"name":"National School of Public Health, Instituto de Salud Carlos III, 28029 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7869-7915","authenticated-orcid":false,"given":"Eugenia","family":"Carrillo","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for Leishmaniasis, National Center for Microbiology, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,16]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2021, February 24). Leishmaniasis Fact Sheet. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/leishmaniasis."},{"key":"ref_2","unstructured":"WHO Expert Committee on the Control of the Leishmaniases & World Health Organization (2021, March 14). Control of the Leishmaniases: Report of a Meeting of the WHO Expert Commitee on the Control of Leishmaniases, Geneva, 22\u201326 March 2010. World Health Organization. Available online: https:\/\/apps.who.int\/iris\/handle\/10665\/44412."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1007\/s00281-020-00788-y","article-title":"Leishmaniasis immunopathology\u2014impact on design and use of vaccines, diagnostics and drugs","volume":"42","author":"Kaye","year":"2020","journal-title":"Semin. Immunopathol."},{"key":"ref_4","unstructured":"World Health Organization (2016). Leishmaniasis in High-Burden Countries: An Epidemiological Update Based on Data Reported in 2014. Wkly Epidemiol. Rec., 91, 287\u2013296. Available online: https:\/\/www.who.int\/wer\/2016\/wer9122.pdf?ua=1."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Verrest, L., Kip, A.E., Musa, A., Schoone, G.J., Schallig, H.D.F.H., Mbui, J., Khalil, E.A.G., Younis, B.M., Olobo, J., and Were, L. (2021). Blood parasite load as an early marker to predict treatment response in visceral leishmaniasis in Eastern Africa. Clin. Infect. Dis., ciab124.","DOI":"10.1093\/cid\/ciab124"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1007\/s40257-015-0114-z","article-title":"Cutaneous Leishmaniasis: Recent Developments in Diagnosis and Management","volume":"16","author":"Reedijk","year":"2015","journal-title":"Am. J. Clin. Dermatol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3147","DOI":"10.1128\/JCM.00372-14","article-title":"Molecular Tools for Diagnosis of Visceral Leishmaniasis: Systematic Review and Meta-Analysis of Diagnostic Test Accuracy","volume":"52","author":"Leeflang","year":"2014","journal-title":"J. Clin. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"E63","DOI":"10.1093\/nar\/28.12.e63","article-title":"Loop-mediated isothermal amplification of DNA","volume":"28","author":"Notomi","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e00386-18","DOI":"10.1128\/JCM.00386-18","article-title":"Development and Evaluation of a Novel Loop-Mediated Isothermal Amplification Assay for Diagnosis of Cutaneous and Visceral Leishmaniasis","volume":"56","author":"Adams","year":"2018","journal-title":"J. Clin. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Mukhtar, M., Ali, S.S., Boshara, S.A., Albertini, A., Monnerat, S., Bessell, P., Mori, Y., Kubota, Y., Ndung\u2019U, J.M., and Cruz, I. (2018). Sensitive and less invasive confirmatory diagnosis of visceral leishmaniasis in Sudan using loop-mediated isothermal amplification (LAMP). PLoS Negl. Trop. Dis., 12.","DOI":"10.1371\/journal.pntd.0006264"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1016\/j.ebiom.2018.10.063","article-title":"Evaluation of point-of-care tests for cutaneous leishmaniasis diagnosis in Kabul, Afghanistan","volume":"37","author":"Vink","year":"2018","journal-title":"EBioMedicine"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Schallig, H.D.F.H., Hu, R.V.P., Kent, A.D., Van Loenen, M., Menting, S., Picado, A., Oosterling, Z., and Cruz, I. (2019). Evaluation of point of care tests for the diagnosis of cutaneous leishmaniasis in Suriname. BMC Infect. Dis., 19.","DOI":"10.1186\/s12879-018-3634-3"},{"key":"ref_13","first-page":"1","article-title":"Evaluation of fluorimetry and direct visualization to interpret results of a loop-mediated isothermal amplification kit to detect Leishmania DNA","volume":"11","author":"Cruz","year":"2018","journal-title":"Parasites Vectors"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"S185","DOI":"10.1016\/S0035-9203(02)90074-X","article-title":"A nested polymerase chain reaction (Ln-PCR) for diagnosing and monitoring Leishmania infantum infection in patients co-infected with human immunodeficiency virus","volume":"96","author":"Cruz","year":"2002","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/j.exppara.2013.03.026","article-title":"An approach for interlaboratory comparison of conventional and real-time PCR assays for diagnosis of human leishmaniasis","volume":"134","author":"Cruz","year":"2013","journal-title":"Exp. Parasitol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2343","DOI":"10.1128\/JCM.02297-05","article-title":"Comparison of New Diagnostic Tools for Management of Pediatric Mediterranean Visceral Leishmaniasis","volume":"44","author":"Cruz","year":"2006","journal-title":"J. Clin. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1111\/j.1365-3164.2009.00824.x","article-title":"Multicentric, controlled clinical study to evaluate effectiveness and safety of miltefosine and allopurinol for canine leishmaniosis","volume":"20","author":"Oliva","year":"2009","journal-title":"Vet. Dermatol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1186\/1756-3305-6-122","article-title":"Characterization of the biology and infectivity of Leishmania infantum viscerotropic and dermotropic strains isolated from HIV+ and HIV- patients in the murine model of visceral leishmaniasis","volume":"6","author":"Cunha","year":"2013","journal-title":"Parasites Vectors"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Cruz, I., Acosta, L., Guti\u00e9rrez, M.N., Nieto, J., Ca\u00f1avate, C., Deschutter, J., and Bornay-Llinares, F.J. (2010). A canine leishmaniasis pilot survey in an emerging focus of visceral leishmaniasis: Posadas (Misiones, Argentina). BMC Infect. Dis., 10.","DOI":"10.1186\/1471-2334-10-342"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.actatropica.2009.09.005","article-title":"PCR for diagnosis and assessment of cure in kala-azar patients in Bangladesh","volume":"113","author":"Salam","year":"2010","journal-title":"Acta Trop."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Alam, M.S., Ghosh, D., Khan, G.M., Islam, M.F., Mondal, D., Itoh, M., Islam, N., and Haque, R. (2011). Survey of domestic cattle for anti-Leishmania antibodies and Leishmania DNA in a visceral leishmaniasis endemic area of Bangladesh. BMC Vet. Res., 7.","DOI":"10.1186\/1746-6148-7-27"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1128\/JCM.05067-11","article-title":"Peripheral Blood Buffy Coat Smear: A Promising Tool for Diagnosis of Visceral Leishmaniasis","volume":"50","author":"Salam","year":"2011","journal-title":"J. Clin. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1590\/S1020-49892010000100012","article-title":"El programa epidat: Usos y perspectivas","volume":"27","author":"Vidal","year":"2010","journal-title":"Revista Panamericana de Salud P\u00fablica"},{"key":"ref_24","unstructured":"Ejov, M., and Dagne, D. (2021, March 14). Strategic Framework for Leishmaniasis Control in the WHO European Region 2014-2020. World Health Organ. Available online: https:\/\/www.euro.who.int\/__data\/assets\/pdf_file\/0017\/245330\/Strategic-framework-for-leishmaniasis-control-in-the-WHO-European-Region-20142020.pdf."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Nzelu, C.O., Kato, H., and Peters, N.C. (2019). Loop-mediated isothermal amplification (LAMP): An advanced molecular point-of-care technique for the detection of Leishmania infection. PLoS Negl. Trop. Dis., 13.","DOI":"10.1371\/journal.pntd.0007698"},{"key":"ref_26","unstructured":"(2021, March 14). WHO Collaborating Centres Global Database [Internet]. Available online: https:\/\/apps.who.int\/whocc\/Detail.aspx?qEjNpHwf6G82WRkvO8QOgA."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/3\/610\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:36:15Z","timestamp":1760160975000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/3\/610"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,16]]},"references-count":26,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2021,3]]}},"alternative-id":["microorganisms9030610"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms9030610","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,16]]}}}