Sabino Raquel, Simões Helena, Veríssimo Cristina
Reference Unit for Parasitic and Fungal Infections, Department of Infectious Diseases, National Institute of Health Dr. Ricardo Jorge, Av. Padre Cruz, 1649-016 Lisbon, Portugal.
J Fungi (Basel). 2020 Jan 10;6(1):11. doi: 10.3390/jof6010011.
Diagnosis of invasive fungal infections is complex, and the lack of standardization of molecular methods is still a challenge. Several methods are available for the diagnosis of invasive aspergillosis, but their effectiveness will depend on the studied population, the patients' comorbidities, and the use of mold active prophylaxis, among others. The ability to determine the identity of the infecting species, and to detect mutations conferring specific resistance patterns directly from DNA extracted from the biological product, is an advantage of nucleic acid testing compared with antigen-based assays. In this study, we to present laboratory cases where the diagnosis of aspergillosis was performed using a real-time multiplex PCR for the detection of DNA in tissue samples, showing its usefulness as one more tool in the diagnosis of aspergillosis in tissue samples. real-time multiplex PCR was also used to detect azole-resistance in some cases. In the majority of the PCR positive cases, cultures remained negative after 60 days. The PCR assay directed to gave positive signals for sensu stricto. Results were confirmed by panfungal PCR, followed by sequencing, revealing 100% homology with sensu stricto. Mutations conferring azole resistance were not detected.
侵袭性真菌感染的诊断很复杂，分子方法缺乏标准化仍是一项挑战。有几种方法可用于诊断侵袭性曲霉病，但其有效性将取决于所研究的人群、患者的合并症以及霉菌活性预防措施的使用等因素。与基于抗原的检测方法相比，核酸检测的一个优势在于能够直接从生物制品中提取的DNA确定感染菌种的身份，并检测赋予特定耐药模式的突变。在本研究中，我们展示了通过实时多重PCR检测组织样本中的DNA来诊断曲霉病的实验室病例，表明其作为组织样本中曲霉病诊断的又一工具的实用性。在某些情况下，实时多重PCR还用于检测唑类耐药性。在大多数PCR阳性病例中，培养60天后仍为阴性。针对烟曲霉狭义种的PCR检测给出了阳性信号。结果通过泛真菌PCR得到证实，随后进行测序，显示与烟曲霉狭义种具有100%的同源性。未检测到赋予唑类耐药性 的突变。
