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Development of a loop-mediated isothermal amplification method for rapid mass-screening of sand flies for Leishmania infection
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) |
URI: | http://hdl.handle.net/2115/54556 |
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)
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Submitter: 加藤 大智
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