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

doi:10.1016/j.actatropica.2015.10.013

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

Research output: Contribution to journal › Article › peer-review

46 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 language	English
Pages (from-to)	116-119
Number of pages	4
Journal	Acta Tropica
Volume	153
DOIs	https://doi.org/10.1016/j.actatropica.2015.10.013
State	Published - 1 Jan 2016

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..

Keywords

FTA
LAMP
Leishmania
Malachite green
Peru

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10.1016/j.actatropica.2015.10.013

Cite this

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, 153, 116-119. https://doi.org/10.1016/j.actatropica.2015.10.013

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title = "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",

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.",

keywords = "FTA, LAMP, Leishmania, Malachite green, Peru",

author = "Nzelu, {Chukwunonso O.} and C{\'a}ceres, {Abraham G.} and Silvia Guerrero-Quincho and Edwin Tineo-Villafuerte and Luis Rodriquez-Delfin and Tatsuyuki Mimori and Hiroshi Uezato and Ken Katakura and Gomez, {Eduardo A.} and Guevara, {Angel G.} and Yoshihisa Hashiguchi and Hirotomo Kato",

note = "Publisher Copyright: {\textcopyright} 2015 Elsevier B.V..",

year = "2016",

month = jan,

day = "1",

doi = "10.1016/j.actatropica.2015.10.013",

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Nzelu, CO, Cáceres, AG, Guerrero-Quincho, S, Tineo-Villafuerte, E, Rodriquez-Delfin, L, Mimori, T, Uezato, H, Katakura, K, Gomez, EA, Guevara, AG, 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, vol. 153, pp. 116-119. https://doi.org/10.1016/j.actatropica.2015.10.013

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. / Nzelu, Chukwunonso O.; Cáceres, Abraham G.; Guerrero-Quincho, Silvia et al.
In: Acta Tropica, Vol. 153, 01.01.2016, p. 116-119.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - 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

AU - Nzelu, Chukwunonso O.

AU - Cáceres, Abraham G.

AU - Guerrero-Quincho, Silvia

AU - Tineo-Villafuerte, Edwin

AU - Rodriquez-Delfin, Luis

AU - Mimori, Tatsuyuki

AU - Uezato, Hiroshi

AU - Katakura, Ken

AU - Gomez, Eduardo A.

AU - Guevara, Angel G.

AU - Hashiguchi, Yoshihisa

AU - Kato, Hirotomo

PY - 2016/1/1

Y1 - 2016/1/1

N2 - 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.

AB - 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.

KW - FTA

KW - LAMP

KW - Leishmania

KW - Malachite green

KW - Peru

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U2 - 10.1016/j.actatropica.2015.10.013

DO - 10.1016/j.actatropica.2015.10.013

M3 - Artículo

C2 - 26516109

AN - SCOPUS:84945354078

SN - 0001-706X

VL - 153

SP - 116

EP - 119

JO - Acta Tropica

JF - Acta Tropica

ER -

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

Abstract

Bibliographical note

Keywords

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