1. 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
- Author
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Angel Guevara, Yoshihisa Hashiguchi, Tatsuyuki Mimori, Luis Rodriquez-Delfin, Edwin Tineo-Villafuerte, Ken Katakura, Silvia Guerrero-Quincho, Abraham G. Cáceres, Hirotomo Kato, Chukwunonso O. Nzelu, Eduardo A. Gomez, and Hiroshi Uezato
- Subjects
Adult ,Male ,0301 basic medicine ,Veterinary (miscellaneous) ,030231 tropical medicine ,Malachite green ,Loop-mediated isothermal amplification ,Leishmaniasis, Cutaneous ,Biology ,Sensitivity and Specificity ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cutaneous leishmaniasis ,LAMP ,Peru ,Rosaniline Dyes ,medicine ,Humans ,Mass Screening ,Colorimetry ,Mass screening ,Leishmania ,Leishmaniasis ,Nucleic acid amplification technique ,medicine.disease ,biology.organism_classification ,Virology ,Molecular biology ,FTA ,030104 developmental biology ,Infectious Diseases ,chemistry ,Insect Science ,Female ,Parasitology ,Nucleic Acid Amplification Techniques - 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 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.
- Published
- 2016