Fungal respiratory tract colonization is a common finding in patients with hematologic neoplasms due to immunosuppression inherent in the diseases and exacerbated by therapy. This greatly increases the risk of fungal infections of the lungs, which is associated with significant mortality. Therefore, reliable diagnostic methods with rapidly available results are needed to administer adequate antifungal therapy. We have established an improved method for fungal DNA extraction and amplification that allows simultaneous detection of fungal families based on a set of multiplexed real-time PCR reactions (fuPCR). We analyzed respiratory rinses and blood of 94 patients with hematological systemic diseases by fuPCR and compared it with the results of culture and serological diagnostic methods. 40 healthy subjects served as controls. Regarding Candida species, the highest prevalence resulted from microbiological culture of respiratory rinses and from detection of antibodies in blood serum in patients (61 and 47%, respectively) and in the control group (29 and 51%, respectively). Detection of other pathogenic yeasts, such as Cryptococcus and Trichosporon, and molds, such as Fusarium, was only possible in patients by fuPCR from both respiratory rinses and whole blood and serum. These fungal species were found statistically significantly more frequent in respiratory rinses collected from patients after myeloablative therapy for stem cell transplantation compared to samples collected before treatment (P < 0.05i). The results show that fuPCR is a valuable complement to culturing and its inclusion in routine mycological diagnostics might be helpful for early detection of pathophysiologically relevant respiratory colonization for patients with hematologic neoplasms.

中文翻译：

---

fuPCR 作为检测稀有真菌病原体的诊断方法，如毛孢菌属、隐球菌属和镰刀菌属

真菌呼吸道定植是血液肿瘤患者的常见发现，这是由于疾病固有的免疫抑制和治疗加剧。这大大增加了肺部真菌感染的风险，这与死亡率显着相关。因此，需要具有快速获得结果的可靠诊断方法来进行适当的抗真菌治疗。我们已经建立了一种改进的真菌 DNA 提取和扩增方法，该方法允许基于一组多重实时 PCR 反应 (fuPCR) 同时检测真菌家族。我们通过 fuPCR 分析了 94 例血液系统疾病患者的呼吸道冲洗液和血液，并将其与培养和血清学诊断方法的结果进行了比较。40 名健康受试者作为对照。关于念珠菌属物种，呼吸道冲洗液的微生物培养和患者（分别为 61% 和 47%）和对照组（分别为 29% 和 51%）血清中抗体检测的患病率最高。其他致病性酵母菌（如隐球菌属和毛孢菌属）和霉菌（如镰刀菌）的检测只能在患者的呼吸道冲洗液、全血和血清中通过 fuPCR 检测。与治疗前收集的样本相比，这些真菌种类在干细胞移植清髓治疗后收集的患者呼吸道冲洗液中的频率显着更高（P < 0.05i）。