School of Medicine and Life Sciences, University of Jinan-Shandong Academy of Medical Sciences, Jinan, China.
State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China.
Sci Rep. 2020 Feb 3;10(1):1640. doi: 10.1038/s41598-020-58245-7.
Early and accurate diagnosis is essential for the targeted management of fungal keratitis (FK), which is one of the major blinding eye diseases worldwide. To elucidate the diagnostic information of high-throughput internal transcribed spacer (ITS) sequencing for identifying causative fungi of FK, 38 patients who were highly suspected of having FK were included in this research. In vivo confocal microscopy, potassium hydroxide smear, and fungal culture were performed to diagnose FK. Culture and ITS sequencing were used to identify causative fungi. We hypothesized that the dominant genus was the result of pathogen identification by ITS sequencing. Thirty-five patients were eventually diagnosed with FK, with fungal pathogens found by confocal microscopy in 27 patients (77.14%), by smear examination in 27 patients (77.14%), by culture in 25 patients (71.43%), and by ITS sequencing in 26 patients (74.29%). Eight causative fungal genera were determined by ITS sequencing, while five causative fungal genera were identified based on the morphology of the cultured pathogens. The results of ITS sequencing and culture were coincident in 10 patients with FK (28.57%). It is concluded that ITS sequencing, to some extent, challenged fungal culture and might be an optional complement in identifying fungal pathogens in corneas.
早期和准确的诊断对于真菌性角膜炎（FK）的靶向治疗至关重要，FK 是全球主要致盲眼病之一。为了阐明高通量内部转录间隔区（ITS）测序在鉴定 FK 致病真菌方面的诊断信息，本研究纳入了 38 名高度疑似 FK 的患者。采用活体共聚焦显微镜、氢氧化钾涂片和真菌培养来诊断 FK，培养和 ITS 测序用于鉴定致病真菌。我们假设优势属是 ITS 测序进行病原体鉴定的结果。最终，35 名患者被诊断为 FK，其中 27 名患者（77.14%）通过共聚焦显微镜、27 名患者（77.14%）通过涂片检查、25 名患者（71.43%）通过培养、26 名患者（74.29%）通过 ITS 测序发现了真菌病原体。ITS 测序确定了 8 个致病真菌属，而根据培养的病原体形态确定了 5 个致病真菌属。10 名 FK 患者的 ITS 测序和培养结果一致（28.57%）。综上所述，ITS 测序在一定程度上挑战了真菌培养，可能是鉴定角膜真菌病原体的一种可选补充方法。
