Contribution of molecular tools for the diagnosis and epidemiology of fungal chronic rhinosinusitis
Contribution of molecular tools for the diagnosis and epidemiology of fungal chronic rhinosinusitis
复制标题
DOI：
10.1093/mmy/myw041
复制
发表时间：
2016-11-01
期刊：
影响因子：
2.9
通讯作者：
Robert-Gangneux, Florence
中科院分区：
文献类型：
--
作者：
Comacle, Pauline;Belaz, Sorya;Robert-Gangneux, Florence
慢性鼻窦炎（CRS）在工业化国家的慢性炎症性疾病中排名第二，是一个重要的公共卫生问题。诊断依赖于一系列的参数，包括临床体征，影像学，组织病理学和真菌学分析窦标本，收集在鼻内窥镜检查。据报道，真菌培养的敏感性很差，即使直接检查是阳性的，因此真菌性慢性鼻窦炎的流行病学是鲜为人知的。本研究评估了在雷恩大学医院3年期间分析的70个连续样本（61例CRS患者）中分子诊断的敏感性。使用靶向ITS 1/ITS 2序列的常规PCR方法进行DNA检测，并对所得扩增产物进行测序。42名患者（69%）被证实患有真菌性CRS，其中只有20名（48%）培养阳性。37/42例（88%）患者被诊断为真菌球，3例为过敏性真菌CRS，2例为未确定的真菌CRS。PCR在所有42例中均为阳性，直接测序允许在除1例外的所有病例中鉴定真菌，并在3例中检测到多重感染。69%的患者中存在烟曲霉; 9.5%存在枝孢样枝孢，Scedosporium sp.，A. nidulans和A.黄色各占7%。在2/19例直接检查阴性的患者中，测序分析显示存在Capnobotryella sp.和C. cladosporoides，在临床环境中与真菌性鼻窦炎相容。总之，ITS 1/ITS 2 PCR的灵敏度是培养的两倍，联合测序提供了准确的真菌CRS流行病学数据。
Chronic rhinosinusitis (CRS) rank second at chronic inflammatory diseases in industrialized countries and are an important public health concern. Diagnosis relies on a set of arguments including clinical signs, imaging, histopathologic and mycological analyses of sinus specimens, collected during nasal endoscopy. The sensitivity of fungal cultures is reported to be poor, even when direct examination is positive, thus the epidemiology of fungal chronic sinusitis is ill-known. This study evaluated the sensitivity of molecular diagnosis in 70 consecutive samples (61 patients with CRS) analysed at the University Hospital of Rennes during a 3-year period. DNA detection was performed using a conventional PCR method targeting the ITS1/ITS2 sequence and the resulting amplification products were sequenced. Fungal CRS was proven in 42 patients (69%), of which only 20 (48%) had a positive culture. 37/42 (88%) patients were diagnosed with a fungus ball, 3 with allergic fungal CRS and 2 with undetermined fungal CRS. PCR was positive in all 42 cases and direct sequencing allowed to identify fungi in all cases but one, and detected multiple infection in 3. Aspergillus fumigatus was present in 69% of patients; Cladosporium cladosporoides in 9.5%, Scedosporium sp., A. nidulans and A. flavus in 7% each. In 2/19 patients with negative direct examination, sequencing analysis revealed the presence of Capnobotryella sp. and C. cladosporoides, in clinical settings compatible with fungal sinusitis. In conclusion, ITS1/ITS2 PCR had a twice better sensitivity than culture, and combined sequencing provides accurate epidemiological data on fungal CRS.