Rapid identification of black grain eumycetoma causative agents using rolling circle amplification

Sarah A Ahmed, Bert Gerrits van den Ende, Ahmed H Fahal, Wendy W J van de Sande, G S de Hoog

    Research output: Contribution to journal/periodicalArticleScientificpeer-review

    35 Citations (Scopus)

    Abstract

    Accurate identification of mycetoma causative agent is a priority for treatment. However, current identification tools are far from being satisfactory for both reliable diagnosis and epidemiological investigations. A rapid, simple, and highly efficient molecular based method for identification of agents of black grain eumycetoma is introduced, aiming to improve diagnostic in endemic areas. Rolling Circle Amplification (RCA) uses species-specific padlock probes and isothermal DNA amplification. The tests were based on ITS sequences and developed for Falciformispora senegalensis, F. tompkinsii, Madurella fahalii, M. mycetomatis, M. pseudomycetomatis, M. tropicana, Medicopsis romeroi, and Trematosphaeria grisea. With the isothermal RCA assay, 62 isolates were successfully identified with 100% specificity and no cross reactivity or false results. The main advantage of this technique is the low-cost, high specificity, and simplicity. In addition, it is highly reproducible and can be performed within a single day.

    Original languageEnglish
    Pages (from-to)e3368
    JournalPLoS Neglected Tropical Diseases
    Volume8
    Issue number12
    DOIs
    Publication statusPublished - Dec 2014

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