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Development of Loop-Mediated Isothermal Amplification (LAMP) Assays for Rapid Detection of Ehrlichia ruminantium

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Title: Development of Loop-Mediated Isothermal Amplification (LAMP) Assays for Rapid Detection of Ehrlichia ruminantium
Authors: Nakao, Ryo Browse this author →KAKEN DB
Stromdahl, Ellen Y. Browse this author
Magona, Joseph W. Browse this author
Faburay, Bonto Browse this author
Namangala, Boniface Browse this author
Malele, Imna Browse this author
Inoue, Noboru Browse this author
Geysen, Dirk Browse this author
Kajino, Kiichi Browse this author →KAKEN DB
Jongejan, Frans Browse this author
Sugimoto, Chihiro Browse this author →KAKEN DB
Issue Date: 19-Nov-2010
Publisher: BioMed Central
Journal Title: BMC Microbiology
Volume: 10
Start Page: 296
Publisher DOI: 10.1186/1471-2180-10-296
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.
Rights: http://creativecommons.org/licenses/by/2.0
Type: article
URI: http://hdl.handle.net/2115/44632
Appears in Collections:人獣共通感染症リサーチセンター (Research Center for Zoonosis Control) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 杉本 千尋

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