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

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 ag...

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Bibliographic Details
Published in:PLoS Neglected Tropical Diseases
Main Authors: Sarah A Ahmed, Bert H G Gerrits van den Ende, Ahmed H Fahal, Wendy W J van de Sande, G S de Hoog
Format: Article in Journal/Newspaper
Language:English
Published: Public Library of Science (PLoS) 2014
Subjects:
Online Access:https://doi.org/10.1371/journal.pntd.0003368
https://doaj.org/article/8a7b200df6654b7c991c547e329e4fa1
Description
Summary: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.