Development of Loop-Mediated Isothermal Amplification (LAMP) assays for rapid detection of Ehrlichia ruminantium

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dc.contributor.author Nakao, Ryo
dc.contributor.author Stromdahl, E.Y.
dc.contributor.author Magona, Joseph W.
dc.contributor.author Faburay, Bonto
dc.contributor.author Namangala, Boniface
dc.contributor.author Malele, Imna
dc.contributor.author Inoue, Noboru
dc.contributor.author Geysen, Dirk
dc.contributor.author Kajino, Kiichi
dc.contributor.author Jongejan, Frans
dc.contributor.author Sugimoto, Chihiro
dc.date.accessioned 2011-04-15T08:03:29Z
dc.date.available 2011-04-15T08:03:29Z
dc.date.issued 2010
dc.description.abstract BACKGROUND: The rickettsial bacterium Ehrlichia ruminantium is the causative agent of heartwater, a potential zoonotic disease of ruminants transmitted by ticks of the genus Amblyomma. The disease is distributed in nearly all of sub-Saharan Africa and some islands of the Caribbean, from where it threatens the American mainland. This report describes the development of two different loop-mediated isothermal amplification (LAMP) assays for sensitive and specific detection of E. ruminantium. RESULTS: Two sets of LAMP primers were designed from the pCS20 and sodB genes. The detection limits for each assay were 10 copies for pCS20 and 5 copies for sodB, which is at least 10 times higher than that of the conventional pCS20 PCR assay. DNA amplification was completed within 60 min. The assays detected 16 different isolates of E. ruminantium from geographically distinct countries as well as two attenuated vaccine isolates. No cross-reaction was observed with genetically related Rickettsiales, including zoonotic Ehrlichia species from the USA. LAMP detected more positive samples than conventional PCR but less than real-time PCR, when tested with field samples collected in sub-Saharan countries. CONCLUSIONS: Due to its simplicity and specificity, LAMP has the potential for use in resource-poor settings and also for active screening of E. ruminantium in both heartwater-endemic areas and regions that are at risk of contracting the disease. en
dc.identifier.citation Nakao, R, Stromdahl, EY, Magona, JW, Faburay, B, Namangala, B, Malele, I, Inoue, N, Geysen, D, Kajino, K, Jongejan, F & Sugimoto, C 2010, ‘Development of Loop-Mediated Isothermal Amplification (LAMP) assays for rapid detection of Ehrlichia ruminantium’, BMC Microbiology, vol. 10, pp. 296-307. [http://www.biomedcentral.com/bmcmicrobiol/] en
dc.identifier.issn 1471-2180
dc.identifier.uri http://hdl.handle.net/2263/16300
dc.language.iso en en
dc.publisher BioMed Central en
dc.relation.requires Adobe Acrobat Reader en
dc.rights © 2010 Nakao et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. en
dc.subject Rickettsial bacterium en
dc.subject Heartwater en
dc.subject Loop-mediated isothermal amplification en
dc.subject LAMP en
dc.subject.lcsh Ehrlichiosis en
dc.subject.lcsh Ruminants -- Diseases en
dc.title Development of Loop-Mediated Isothermal Amplification (LAMP) assays for rapid detection of Ehrlichia ruminantium en
dc.type Article en


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