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Development of a loop-mediated isothermal amplification method for rapid mass-screening of sand flies for Leishmania infection

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Title: Development of a loop-mediated isothermal amplification method for rapid mass-screening of sand flies for Leishmania infection
Authors: Nzelu, Chukwunonso O. Browse this author
Gomez, Eduardo A. Browse this author
Cáceres, Abraham G. Browse this author
Sakurai, Tatsuya Browse this author
Martini-Robles, Luiggi Browse this author
Uezato, Hiroshi Browse this author →KAKEN DB
Mimori, Tatsuyuki Browse this author →KAKEN DB
Katakura, Ken Browse this author →KAKEN DB
Hashiguchi, Yoshihisa Browse this author →KAKEN DB
Kato, Hirotomo Browse this author →KAKEN DB
Keywords: Leishmania
Sand fly
Malachite green
Loop-mediated isothermal amplification (LAMP)
Issue Date: Apr-2014
Publisher: Elsevier
Journal Title: Acta Tropica
Volume: 132
Start Page: 1
End Page: 6
Publisher DOI: 10.1016/j.actatropica.2013.12.016
PMID: 24388795
Abstract: Entomological monitoring of Leishmania infection in leishmaniasis endemic areas offers epidemiologic advantages for predicting the risk and expansion of the disease, as well as evaluation of the effectiveness of control programs. In this study, we developed a highly sensitive loop-mediated isothermal amplification (LAMP) method for the mass screening of sand flies for Leishmania infection based on the 18S rRNA gene. The LAMP technique could detect 0.01 parasites, which was more sensitive than classical PCR. The method was robust and could amplify the target DNA within 1 h from a crude sand fly template without DNA purification. Amplicon detection could be accomplished by the newly developed colorimetric malachite green (MG)—mediated naked eye visualization. Pre-addition of MG to the LAMP reaction solution did not inhibit amplification efficiency. The field applicability of the colorimetric MG-based LAMP assay was demonstrated with 397 field-caught samples from the endemic areas of Ecuador and eight positive sand flies were detected. The robustness, superior sensitivity, and ability to produce better visual discriminatory reaction products than existing LAMP fluorescence and turbidity assays indicated the field potential usefulness of this new method for surveillance and epidemiological studies of leishmaniasis in developing countries.
Type: article (author version)
Appears in Collections:獣医学院・獣医学研究院 (Graduate School of Veterinary Medicine / Faculty of Veterinary Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 加藤 大智

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