Internal transcribed spacer (ITS) sequencing for Acute Lung Injury Registry

In critically ill patients, variation in the host inflammatory responses have been associated with poor outcomes. However, the biological mechanisms underlying this heterogeneity have not yet been defined. We investigated whether variation fungal communities (mycobiome) in the respiratory tract are associated with host inflammation and innate immune system activation and clinical and patient centered outcomes.We collected endotracheal aspirates (ETA), oral wash and plasma samples from critically ill patients. We used extracted DNA from ETAs and performed fungal rRNA gene sequencing (internal transcribed spacer [ITS]) on the Illumina MiSeq platform to characterize the lower respiratory tract mycobiome. We derived diversity metrics to classify patients into strata based on the alpha diversity of the mycobiome and examined for associations with diversity strata and clinical and patient centered outcomes.

针对重症患者而言，宿主炎症反应的异质性与不良临床结局存在相关性。然而，该异质性背后的生物学机制尚未阐明。本研究旨在探讨呼吸道真菌群落（mycobiome，真菌组）的异质性是否与宿主炎症反应、先天免疫系统激活以及临床结局和以患者为中心的结局相关。我们从重症患者中收集了气管吸出物（endotracheal aspirates, ETA）、口腔冲洗液与血浆样本。我们提取气管吸出物中的DNA，并依托Illumina MiSeq测序平台对真菌核糖体RNA基因的内部转录间隔区（internal transcribed spacer, ITS）进行测序，以此对下呼吸道真菌组进行特征分析。我们基于真菌组的α多样性计算多样性指标，将患者划分为不同分层，并分析该多样性分层与临床结局及以患者为中心的结局之间的相关性。