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

• Published in: Acta tropica 2016-01, Vol.153, p.116-119
• Main Authors: Nzelu, Chukwunonso O., Cáceres, Abraham G., Guerrero-Quincho, Silvia, Tineo-Villafuerte, Edwin, Rodriquez-Delfin, Luis, Mimori, Tatsuyuki, Uezato, Hiroshi, Katakura, Ken, Gomez, Eduardo A., Guevara, Angel G., Hashiguchi, Yoshihisa, Kato, Hirotomo
• Format: Article
• Language: English
• Subjects: Adult; Colorimetry - methods; Female; FTA; Humans; LAMP; Leishmania; Leishmania - isolation & purification; Leishmaniasis, Cutaneous - diagnosis; Malachite green; Male; Mass Screening - methods; Nucleic Acid Amplification Techniques; Peru; Rosaniline Dyes - analysis; Sensitivity and Specificity
• ISSN: 0001-706X; 1873-6254
• DOI: 10.1016/j.actatropica.2015.10.013

Colorimetric malachite green based FTA-LAMP assay for cutaneous leishmaniasis. [Display omitted] •A LAMP assay for rapid detection of Leishmania DNA from an FTA card was established.•The colorimetric FTA-LAMP was highly sensitive enough to detect 0.01 parasites.•The field applicability was demonstrated with clinical samples collected from patients with CL in Peru.•The FTA-LAMP will be an important tool for diagnosis of cutaneous leishmaniasis. 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 60min 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.