A rapid molecular diagnosis of cutaneous leishmaniasis by colorimetric malachite green-loop-mediated isothermal amplification (LAMP) combined with an FTA card as a direct sampling tool

Chukwunonso O. Nzelu, Abraham G. Cáceres, Silvia Guerrero-Quincho, Edwin Tineo-Villafuerte, Luis Rodriquez-Delfin, Tatsuyuki Mimori, Hiroshi Uezato, Ken Katakura, Eduardo A. Gomez, Angel G. Guevara, Yoshihisa Hashiguchi, Hirotomo Kato

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22 Scopus citations

Abstract

© 2015 Elsevier B.V.. Leishmaniasis 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. °C for 60. min 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 μl. 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.
Original languageAmerican English
Pages (from-to)116-119
Number of pages4
JournalActa Tropica
DOIs
StatePublished - 1 Jan 2016

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    Nzelu, C. O., Cáceres, A. G., Guerrero-Quincho, S., Tineo-Villafuerte, E., Rodriquez-Delfin, L., Mimori, T., Uezato, H., Katakura, K., Gomez, E. A., Guevara, A. G., Hashiguchi, Y., & Kato, H. (2016). A rapid molecular diagnosis of cutaneous leishmaniasis by colorimetric malachite green-loop-mediated isothermal amplification (LAMP) combined with an FTA card as a direct sampling tool. Acta Tropica, 116-119. https://doi.org/10.1016/j.actatropica.2015.10.013