Ahmed Sarah A, van den Ende Bert H G Gerrits, Fahal Ahmed H, van de Sande Wendy W J, de Hoog G S
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.
CBS-KNAW Fungal Biodiversity Centre, Utrecht, The Netherlands.
PLoS Negl Trop Dis. 2014 Dec 4;8(12):e3368. doi: 10.1371/journal.pntd.0003368. eCollection 2014 Dec.
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.
准确鉴定足菌肿病原体是治疗的首要任务。然而，目前的鉴定工具在可靠诊断和流行病学调查方面都远不能令人满意。本文介绍了一种快速、简单且高效的基于分子的黑粒真菌性足菌肿病原体鉴定方法，旨在改善流行地区的诊断。滚环扩增（RCA）使用物种特异性锁式探针和等温DNA扩增。这些测试基于ITS序列，针对塞内加尔镰孢菌、汤普金斯镰孢菌、法哈利马杜拉菌、马杜拉足菌肿马杜拉菌、假马杜拉足菌肿马杜拉菌、热带马杜拉菌、罗梅罗医学拟青霉和灰色 Trematosphaeria 菌开发。通过等温RCA检测，成功鉴定了62株分离株，特异性为100%，无交叉反应或假结果。该技术的主要优点是成本低、特异性高且操作简单。此外，它具有高度可重复性，可在一天内完成。
