title: Rapid identification of black grain eumycetoma causative agents using rolling circle amplification.
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.
authors.affiliation: Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan; CBS-KNAW Fungal Biodiversity Centre, Utrecht, The Netherlands; Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Ahmed
authors.affiliation: CBS-KNAW Fungal Biodiversity Centre, Utrecht, The Netherlands.
authors.forename: Bert H G Gerrits
authors.initials: BH
authors.lastname: van den Ende
authors.affiliation: Mycetoma Research Centre, University of Khartoum, Khartoum, Sudan.
authors.forename: Ahmed H
authors.initials: AH
authors.lastname: Fahal
authors.affiliation: Department of Medical Microbiology and Infectious Diseases, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Wendy W J
authors.initials: WW
authors.lastname: van de Sande
authors.affiliation: Peking University Health Science Center, Research Center for Medical Mycology, Beijing, China; Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, China; Shanghai Institute of Medical Mycology, Changzheng Hospital, Second Military Medical University, Shanghai, China; Basic Pathology Department, Federal University of Paraná State, Curitiba, Brazil; King Abdulaziz University, Jeddah, Saudi Arabia.
authors.forename: G S
authors.initials: GS
authors.lastname: de Hoog
mesh_terms: D006801:Humans; D020461:Madurella / Q000235:genetics*; D008271:Mycetoma / Q000382:microbiology*; D016533:Mycological Typing Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D014871:Water Microbiology