Microbiology, Parasitology and Pathology Post-Graduation Program, Department of Pathology, Federal University of Paraná, Curitiba, Brazil.
Engineering Bioprocess and Biotechnology Post-Graduation Program, Department of Bioprocess Engineering and Biotechnology, Federal University of Paraná, Curitiba, Brazil.
Mycopathologia. 2019 Aug;184(4):493-504. doi: 10.1007/s11046-019-00359-2. Epub 2019 Jul 17.
The species belonging to the genus Fonsecaea are the main causative agents of chromoblastomycosis. The invasive potential of Fonsecaea differs significantly among its various sibling species. Moreover, the lack of clarity on the virulence and availability of precise markers to distinguish and detect Fonsecaea species is attributed to the different ways of dissemination and pathogenicity. Therefore, the present study aimed to propose new molecular tools to differentiate between sibling species causing chromoblastomycosis. We used an infection model of chromoblastomycosis in BALB/c to study species-specific molecular markers for the in vivo detection of Fonsecaea species in biological samples. Specific primers based on the CBF5 gene were developed for Fonsecaea pedrosoi, Fonsecaea monophora, Fonsecaea nubica, and Fonsecaea pugnacius. In addition, a padlock probe was designed for F. pugnacius based on ITS sequences. We also assessed the specificity of Fonsecaea species using in silico, in vitro, and in vivo assays. The results showed that markers and probes could effectively discriminate the species in both clinical and environmental samples, enabling bioprospecting of agents of chromoblastomycosis, thereby elucidating the infection route of the disease.
属于绵霉属的物种是着色芽生菌病的主要病原体。绵霉属的各种姊妹种之间的侵袭潜力有很大差异。此外，由于对毒力和区分及检测绵霉属物种的精确标记物的可用性缺乏清晰认识，导致了不同的传播方式和致病性。因此，本研究旨在提出新的分子工具来区分引起着色芽生菌病的姊妹种。我们使用 BALB/c 动物的着色芽生菌病感染模型来研究用于体内检测生物样本中绵霉属物种的种特异性分子标记物。基于 CBF5 基因开发了针对佩德罗索绵霉、单头绵霉、努比亚绵霉和多形绵霉的特异性引物。此外，还基于 ITS 序列为 F. pugnacius 设计了一个套锁探针。我们还使用计算机模拟、体外和体内检测评估了绵霉属物种的特异性。结果表明，标记物和探针可以有效地在临床和环境样本中区分这些物种，从而进行着色芽生菌病病原体的生物勘探，阐明疾病的感染途径。
