The genus Madurella comprising four species, M. fahalii, M. mycetomatis, M. pseudomycetomatis, and M. tropicana, represents the prevalent cause of eumycetoma worldwide. The four species are phenotypically similar and cause an invariable clinical picture, but differ markedly in their susceptibility to antifungal drugs, and epidemiological pattern. Therefore, specific identification is required for optimal management of Madurella infection and to reveal proper epidemiology of the species. In this study, a novel multiplex real-time PCR targeting the four Madurella species was developed and standardized. Evaluation of the assay using reference strains of the target and non-target species resulted in 100% specificity, high analytical reproducibility (R2 values >0.99) and a lowest detection limit of 3 pg target DNA. The accuracy of the real-time PCR was further assessed using biopsies from eumycetoma suspected patients. Unlike culture and DNA sequencing as gold standard diagnostic methods, the real-time PCR yielded accurate diagnosis with specific identification of the causative species in three hours compared to one or two weeks required for culture. The novel method reduces turnaround time as well as labor intensity and high costs associated with current reference methods.

中文翻译：

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Madurella实时荧光定量PCR，这是一种用于诊断真菌样瘤的新方法。

马杜氏杆菌属包括四个种类，即法氏梭状芽胞杆菌，食肉支原体，假支原体支原体和热带支原体，这是世界范围内杜仲的普遍原因。这四个种类在表型上相似，并产生不变的临床图像，但在抗真菌药的敏感性和流行病学模式上有明显不同。因此，需要进行特殊鉴定才能对马氏杆菌感染进行最佳处理，并揭示该物种的适当流行病学。在这项研究中，开发并标准化了针对四种马杜氏菌的新型多重实时PCR。使用靶标和非靶标物种的参考菌株对测定进行评估，结果得出100％的特异性，高分析重现性（R2值> 0.99）和最低检测限3 pg靶DNA。实时PCR的准确性通过使用疑似真菌病患者的活检进一步评估。与作为金标准诊断方法的培养和DNA测序不同，实时PCR产生的准确诊断具有对致病菌种的特​​异性鉴定，而培养所需的时间为一到两周，因此可以在三小时内进行特异性鉴定。该新颖方法减少了周转时间以及与当前参考方法相关的劳动强度和高成本。