Wang James C, Bergeron Mathieu, Andersen Heidi, Tikhtman Raisa, Haslam David, Hunter Tammy, Herr Andrew B, de Alarcon Alessandro
Department of Otolaryngology-Head and Neck Surgery, Cincinnati, OH, U.S.A.
Division of Pediatric Otolaryngology-Head and Neck Surgery, Cincinnati, OH, U.S.A.
Laryngoscope. 2019 Feb;129(2):317-323. doi: 10.1002/lary.27356. Epub 2018 Nov 22.
Biofilm formation on medical devices such as tracheostomy tubes (TTs) is a serious problem. The clinical impact of biofilms on the airway is still unclear. Biofilms may play a role in granulation tissue development, recurrent airway infections, and failure of laryngotracheal reconstructions. The microbial ecology on TTs has yet to be elucidated. The purpose of this study was to determine the feasibility of shotgun metagenomics to assess the biodistribution of microorganisms on TTs.
Four TTs were collected from pediatric patients (1.4-10.2 years) with (n = 2) and without (n = 2) granulation tissue formation. Duration of TT placement prior to retrieval from patients ranged from 5 to 365 days. DNA extraction was performed using the MO BIO UltraClean Microbial Isolation (Mo Bio Laboratories, Carlsbad, CA). Library generation using Nextera XT adapters (Illumina Inc., San Diego, CA) and metagenomic shotgun sequencing was performed using the Illumina NextSeq500 (Illumina Inc, San Diego, CA). Salinibacter ruber, a species not found in mammalian microbiome communities, was used as a DNA standard and represented 0.7% to 5.7% of the microbiome, ensuring good quality and abundance of sample DNA.
Metagenomic shotgun sequencing was successful for all patients. In TTs associated with granuloma, Fusobacterium nucleatum, Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae were predominant, most of which are considered pathogens. From TTs without granulomas, Neisseria mucosa, Neisseria sicca, Acinetobacter baumannii, and Haemophilus parainfluenzae were identified, primarily consistent with respiratory microbiome.
This study reveals that metagenomic shotgun sequencing of biofilms formed on pediatric TTs is feasible with an apparent difference in microbiome for patients with granulation tissue. Further studies are necessary to elucidate the pathogenesis of microbial ecology and its role in airway disease in patients with TTs.
2c Laryngoscope, 129:317-323, 2019.
在诸如气管造口管（TTs）等医疗器械上形成生物膜是一个严重问题。生物膜对气道的临床影响仍不明确。生物膜可能在肉芽组织形成、反复气道感染以及喉气管重建失败中起作用。TTs上的微生物生态尚未阐明。本研究的目的是确定鸟枪法宏基因组学评估TTs上微生物生物分布的可行性。
从患有（n = 2）和未患有（n = 2）肉芽组织形成的儿科患者（1.4 - 10.2岁）中收集4根TTs。从患者体内取出TTs之前的放置时间为5至365天。使用MO BIO UltraClean Microbial Isolation（Mo Bio Laboratories，卡尔斯巴德，加利福尼亚州）进行DNA提取。使用Nextera XT接头（Illumina Inc.，圣地亚哥，加利福尼亚州）进行文库构建，并使用Illumina NextSeq500（Illumina Inc，圣地亚哥，加利福尼亚州）进行宏基因组鸟枪法测序。盐红菌是一种在哺乳动物微生物群落中未发现的物种，用作DNA标准品，占微生物组的0.7%至5.7%，确保样本DNA的质量和丰度良好。
对所有患者进行鸟枪法宏基因组测序均成功。在与肉芽肿相关的TTs中，具核梭杆菌、流感嗜血杆菌、卡他莫拉菌和肺炎链球菌占主导，其中大多数被认为是病原体。在无肉芽肿的TTs中，鉴定出黏膜奈瑟菌、微黄奈瑟菌、鲍曼不动杆菌和副流感嗜血杆菌，主要与呼吸道微生物组一致。
本研究表明，对儿科TTs上形成的生物膜进行鸟枪法宏基因组测序是可行的，肉芽组织患者的微生物组存在明显差异。需要进一步研究以阐明微生物生态的发病机制及其在TTs患者气道疾病中的作用。
2c 《喉镜》，2019年，第129卷，第317 - 323页
