Zhao Yi Chen, Bassiouni Ahmed, Tanjararak Kangsadarn, Vreugde Sarah, Wormald Peter-John, Psaltis Alkis James
Department of Surgery-Otorhinolaryngology, Head and Neck Surgery, University of Adelaide, Adelaide, Australia.
Laryngoscope. 2018 Jan;128(1):16-22. doi: 10.1002/lary.26702. Epub 2017 Jul 4.
Next-generation sequencing increases the sensitivity of fungal identification and may improve our understanding of the role that fungi play in sinus health and disease, which remains incompletely understood. We sequenced the internal transcribed spacer (ITS) amplicon to explore the role of the mycobiome in chronic rhinosinusitis (CRS).
Swabs were collected intraoperatively from the middle meatus of 90 patients (63 with CRS; 27 controls). DNA was extracted, and ITS amplicon concentration was measured using fluorometry. Internal transcribed spacer amplicons were sequenced on the Illumina MiSeq (Illumina Inc San Diego CA). Sequencing data were analyzed using QIIME.
Using conventional detection techniques of culture and histology, fungi only were identified in nine of 63 (14.3%) CRS patients (fungus-identified group); the remaining 54 CRS patients and all controls did not have fungus identified using the traditional techniques. This fungus-identified group had a significantly higher average ITS concentration and a significantly lower Shannon's diversity index compared to the other two groups. The most abundant organism sequenced was Aspergillus (35.22% of all sequences). Multivariate analysis showed that positive fungal detection using traditional techniques and computed tomography (CT) double densities were the most important clinical predictors of a high fungal biomass, whereas Lund-Mackay score, polyps, eosinophilia, and eosinophilic mucus were not significant in comparison.
Fungal biomass estimated through ITS amplicon concentration correlated with traditional fungal detection techniques and CT double densities. Our results suggest that fungal dysbiosis only occurs in the sinuses of a selected subset of patients, and therefore could not be a universal determinant of sinus disease pathogenesis in all CRS patients.
NA. Laryngoscope, 128:16-22, 2018.
新一代测序提高了真菌鉴定的敏感性，可能有助于增进我们对真菌在鼻窦健康与疾病中所起作用的理解，而目前对此的理解仍不完整。我们对内部转录间隔区（ITS）扩增子进行测序，以探究真菌微生物群在慢性鼻-鼻窦炎（CRS）中的作用。
术中从90例患者（63例CRS患者；27例对照）的中鼻道采集拭子。提取DNA，并使用荧光法测量ITS扩增子浓度。在Illumina MiSeq（Illumina公司，加利福尼亚州圣地亚哥）上对内部转录间隔区扩增子进行测序。使用QIIME分析测序数据。
使用传统的培养和组织学检测技术，仅在63例CRS患者中的9例（14.3%）中鉴定出真菌（真菌鉴定组）；其余54例CRS患者和所有对照使用传统技术均未鉴定出真菌。与其他两组相比，该真菌鉴定组的平均ITS浓度显著更高，香农多样性指数显著更低。测序出的最丰富的生物体是曲霉（占所有序列的35.22%）。多变量分析显示，使用传统技术进行的真菌阳性检测和计算机断层扫描（CT）双密度是真菌生物量高的最重要临床预测因素，而相比之下，Lund-Mackay评分、息肉、嗜酸性粒细胞增多和嗜酸性黏液并不显著。
通过ITS扩增子浓度估算的真菌生物量与传统真菌检测技术及CT双密度相关。我们的结果表明，真菌生态失调仅发生在部分特定患者的鼻窦中，因此不可能是所有CRS患者鼻窦疾病发病机制的普遍决定因素。
无。《喉镜》，2018年，第128卷，第16 - 22页。
