Cytoimmunodiagnostic Laboratory, Department of Public Health and Pediatrics, University of Turin, Piazza Polonia 94, 10126 Turin, Italy.
Bacteriology and Mycology Laboratory, Department of Public Health and Pediatrics, University of Turin, Piazza Polonia 94, 10126 Turin, Italy.
Int J Mol Sci. 2022 Dec 1;23(23):15066. doi: 10.3390/ijms232315066.
Infections triggered by filamentous fungi placed in the order Mucorales, phylum Zygomycota, can cause serious harm to immunocompromised patients. Since there is lack of a standardized PCR (polymerase chain reaction) assay for early diagnosis of this fungal infection, this work was aimed to develop a new PCR assay able to detect the presence of Mucorales genera in clinical specimens. Here, we describe a novel diagnostic TaqMan MGB probe assay for precise and rapid detection of the most common clinical species of Mucorales. Zygomycete-specific oligonucleotides were designed to specifically amplify and bind highly conserved sequences of fungal 28S rRNA gene. Additionally, we succeeded in differentiating Mucorales species (i.e., , , , and ) in artificially infected serum samples, suggesting that the quantitative capability of this real-time PCR assay could potentially optimize the diagnosis of mucormycosis.
在丝孢真菌目中的毛霉目真菌(Mucorales)、接合菌门(Zygomycota)的感染会对免疫功能低下的患者造成严重伤害。由于缺乏用于早期诊断这种真菌感染的标准化聚合酶链反应(PCR)检测方法,本研究旨在开发一种新的 PCR 检测方法,能够检测临床标本中毛霉目属的存在。在这里,我们描述了一种新型的诊断性 TaqMan MGB 探针检测方法,用于精确和快速检测最常见的临床毛霉目真菌。设计了接合菌属特异性寡核苷酸,以特异性扩增和结合真菌 28S rRNA 基因的高度保守序列。此外,我们成功地在人工感染的血清样本中区分了毛霉目真菌(即 , , , ),这表明该实时 PCR 检测方法的定量能力有可能优化毛霉病的诊断。