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 German Caceres Lazaro, 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

Research output: Contribution to journalArticlepeer-review

29 Scopus citations

Abstract

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 languageEnglish
Pages (from-to)116-119
Number of pages4
JournalActa Tropica
Volume153
DOIs
StatePublished - 1 Jan 2016
Externally publishedYes

Bibliographical note

Funding Information:
The authors thanks the patients for giving consent to be a part of the study and all medical doctors and health staff at the different centers in Peru for their active participation during sampling. This study was financially supported by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan (Grant nos. 25257501 and 23256002 ), and the Program for Leading Graduate Schools “Fostering Global Leaders in Veterinary Science for Contributing to One Health” (FO1), MEXT, Japan. CON is a recipient of the Japanese Government Ministry of Education, Culture, Sports, Science and Technology (MEXT) Ph.D. fellowship.

Publisher Copyright:
© 2015 Elsevier B.V..

Copyright:
Copyright 2015 Elsevier B.V., All rights reserved.

Keywords

  • FTA
  • LAMP
  • Leishmania
  • Malachite green
  • Peru

Fingerprint Dive into the research topics of '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'. Together they form a unique fingerprint.

Cite this