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

R Nakao, EY Stromdahl, JW Magona, B Faburay, B Namangala, I Malele, N Inoue, D Geysen, K Kajino, F Jongejan, C Sugimoto

    Research output: Contribution to journalA1: Web of Science-article

    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
    Original languageEnglish
    JournalBMC Microbiology
    Volume10
    Pages (from-to)296
    Number of pages11
    ISSN1471-2180
    DOIs
    Publication statusPublished - 2010

    Keywords

    • B780-tropical-medicine
    • Rickettsial diseases
    • Ehrlichia ruminantium
    • Heartwater
    • Zoonoses
    • Ticks
    • Amblyomma variegatum
    • Amblyomma americanum
    • Detection
    • Rapid diagnostic tests
    • Amplification
    • Primers
    • Assays
    • Identification
    • Comparison
    • Polymerase chain reaction
    • PCR
    • Real-time
    • Sensitivity
    • Specificity
    • Cross reactions
    • America-Latin
    • Caribbean
    • Africa-General

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