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患有和未患有囊性纤维化的儿童下呼吸道中真菌的检测与鉴定。

Detection and identification of fungi in the lower airway of children with and without cystic fibrosis.

作者信息

O'Connor John B, Wagner Brandie D, Harris J Kirk, Frank Daniel N, Clabots Diana E, Laguna Theresa A

机构信息

Division of Pulmonary and Sleep Medicine, Ann & Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, United States.

University of Colorado School of Medicine, Aurora, CO, United States.

出版信息

Front Microbiol. 2023 Feb 9;14:1119703. doi: 10.3389/fmicb.2023.1119703. eCollection 2023.

Abstract

INTRODUCTION

Airway infection and inflammation lead to the progression of obstructive lung disease in persons with cystic fibrosis (PWCF). However, cystic fibrosis (CF) fungal communities, known drivers of CF pathophysiology, remain poorly understood due to the shortcomings of traditional fungal culture. Our objective was to apply a novel small subunit rRNA gene (SSU-rRNA) sequencing approach to characterize the lower airway mycobiome in children with and without CF.

METHODS

Bronchoalveolar lavage fluid (BALF) samples and relevant clinical data were collected from pediatric PWCF and disease control (DC) subjects. Total fungal load (TFL) was measured using quantitative PCR, and SSU-rRNA sequencing was used for mycobiome characterization. Results were compared across groups, and Morisita-Horn clustering was performed.

RESULTS

161 (84%) of the BALF samples collected had sufficient load for SSU-rRNA sequencing, with amplification being more common in PWCF. BALF from PWCF had increased TFL and increased neutrophilic inflammation compared to DC subjects. PWCF exhibited increased abundance of and , while , , and Pleosporales were prevalent in both groups. CF and DC samples showed no clear differences in clustering when compared to each other or to negative controls. SSU-rRNA sequencing was used to profile the mycobiome in pediatric PWCF and DC subjects. Notable differences were observed between the groups, including the abundance of and .

DISCUSSION

Fungal DNA detected in the airway could represent a combination of pathogenic fungi and environmental exposure (e.g., dust) to fungus indicative of a common background signature. Next steps will require comparisons to airway bacterial communities.

摘要

引言

气道感染和炎症会导致囊性纤维化患者(PWCF)的阻塞性肺病进展。然而,由于传统真菌培养的局限性,囊性纤维化(CF)真菌群落作为CF病理生理学的已知驱动因素,仍未得到充分了解。我们的目标是应用一种新型的小亚基核糖体RNA基因(SSU-rRNA)测序方法,来表征有和没有CF的儿童的下呼吸道真菌群落。

方法

从儿科PWCF和疾病对照(DC)受试者中收集支气管肺泡灌洗(BALF)样本和相关临床数据。使用定量PCR测量总真菌负荷(TFL),并使用SSU-rRNA测序进行真菌群落表征。对各组结果进行比较,并进行森下-霍恩聚类分析。

结果

收集的161份(84%)BALF样本有足够的负荷用于SSU-rRNA测序,扩增在PWCF中更常见。与DC受试者相比,PWCF的BALF的TFL增加,中性粒细胞炎症增加。PWCF中 和 的丰度增加,而 、 和格孢腔菌目在两组中都很普遍。与阴性对照相比,CF和DC样本在聚类方面没有明显差异。SSU-rRNA测序用于分析儿科PWCF和DC受试者的真菌群落。在两组之间观察到显著差异,包括 和 的丰度。

讨论

在气道中检测到的真菌DNA可能代表致病真菌和环境中真菌暴露(如灰尘)的组合,表明存在共同的背景特征。下一步需要与气道细菌群落进行比较。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/75fa/9948248/90288ca1a3c9/fmicb-14-1119703-g001.jpg

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