2024-03-28T15:19:19Zhttps://eprints.lib.hokudai.ac.jp/dspace-oai/requestoai:eprints.lib.hokudai.ac.jp:2115/639832022-11-17T02:08:08Zhdl_2115_20048hdl_2115_140A rapid molecular diagnosis of cutaneous leishmaniasis by colorimetric malachite green-loop-mediated isothermal amplification (LAMP) combined with an FTA card as a direct sampling toolNzelu, Chukwunonso O.Caceres, Abraham G.Guerrero-Quincho, SilviaTineo-Villafuerte, EdwinRodriquez-Delfin, LuisMimori, TatsuyukiUezato, HiroshiKatakura, KenGomez, Eduardo A.Guevara, Angel G.Hashiguchi, YoshihisaKato, HirotomoLeishmaniaLAMPFTAMalachite greenPeru491Leishmaniasis remains one of the world's most neglected diseases, and early detection of the infectious agent, especially in developing countries, will require a simple and rapid test. In this study, we established a quick, one-step, single-tube, highly sensitive loop-mediated isothermal amplification (LAMP) assay for rapid detection of Leishmania DNA from tissue materials spotted on an FTA card. An FTA-LAMP with pre-added malachite green was performed at 64 degrees C for 60 mm using a heating block and/or water bath and DNA amplification was detected immediately after incubation. The LAMP assay had high detection sensitivity down to a level of 0.01 parasites per mu l. The field- and clinic-applicability of the colorimetric FTA-LAMP assay was demonstrated with 122 clinical samples collected from patients suspected of having cutaneous leishmaniasis in Peru, from which 71 positives were detected. The LAMP assay in combination with an FTA card described here is rapid and sensitive, as well as simple to perform, and has great potential usefulness for diagnosis and surveillance of leishmaniasis in endemic areas. (C) 2015 Elsevier B.V. All rights reserved.ElsevierJournal Articleapplication/pdfhttp://hdl.handle.net/2115/63983https://eprints.lib.hokudai.ac.jp/dspace/bitstream/2115/63983/1/ActaTropica_v.153%20p.116-119.pdf0001-706X1873-6254AA00510523Acta Tropica1531161192016-01enginfo:pmid/26516109info:doi/10.1016/j.actatropica.2015.10.013©2016. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/author