Department of Surgery, University of Texas Southwestern Medical Center, Dallas, Texas, United States of America.
Department of Microbiology, Immunology, and Genetics, University of North Texas Health Science Center, Fort Worth, Texas, United States of America.
PLoS One. 2018 Oct 24;13(10):e0205788. doi: 10.1371/journal.pone.0205788. eCollection 2018.
Mechanically ventilated surgical patients have a variety of bacterial flora that are often undetectable by traditional culture methods. The source of infection in many of these patients remains unclear. To address this clinical problem, the microbiome profile and host inflammatory response in bronchoalveolar lavage samples from the surgical intensive care unit were examined relative to clinical pathology diagnoses. The hypothesis was tested that clinical diagnosis of respiratory tract flora were similar to culture positive lavage samples in both microbiome and inflammatory profile. Bronchoalveolar lavage samples were collected in the surgical intensive care unit as standard of care for intubated individuals with a clinical pulmonary infection score of >6 or who were expected to be intubated for >48 hours. Cytokine analysis was conducted with the Bioplex Pro Human Th17 cytokine panel. The microbiome of the samples was sequenced for the 16S rRNA region using the Ion Torrent. Microbiome diversity analysis showed the culture-positive samples had the lowest levels of diversity and culture negative with the highest based upon the Shannon-Wiener index (culture positive: 0.77 ± 0.36, respiratory tract flora: 2.06 ± 0.73, culture negative: 3.97 ± 0.65). Culture-negative samples were not dominated by a single bacterial genera. Lavages classified as respiratory tract flora were more similar to the culture-positive in the microbiome profile. A comparison of cytokine expression between groups showed increased levels of cytokines (IFN-g, IL-17F, IL-1B, IL-31, TNF-a) in culture-positive and respiratory tract flora groups. Culture-positive samples exhibited a more robust immune response and reduced diversity of bacterial genera. Lower cytokine levels in culture-negative samples, despite a greater number of bacterial species, suggest a resident nonpathogenic bacterial community may be indicative of a normal pulmonary environment. Respiratory tract flora samples were most similar to the culture-positive samples and may warrant classification as culture-positive when considering clinical treatment.
机械通气的外科患者有多种细菌菌群,这些菌群通常无法通过传统的培养方法检测到。这些患者的感染源仍不清楚。为了解决这一临床问题,研究人员检查了外科重症监护病房支气管肺泡灌洗液样本的微生物组谱和宿主炎症反应与临床病理诊断的关系。研究假设是,呼吸道菌群的临床诊断与培养阳性的灌洗液样本在微生物组和炎症谱方面相似。支气管肺泡灌洗液样本是作为标准护理程序在外科重症监护病房中采集的,采集对象为临床肺部感染评分(CPIS)>6 分或预计需插管>48 小时的插管患者。细胞因子分析采用 Bioplex Pro 人类 Th17 细胞因子面板进行。使用 Ion Torrent 对样本的 16S rRNA 区域进行微生物组测序。微生物组多样性分析显示,培养阳性样本的多样性最低,培养阴性样本的多样性最高,基于 Shannon-Wiener 指数(培养阳性:0.77±0.36,呼吸道菌群:2.06±0.73,培养阴性:3.97±0.65)。培养阴性样本没有被单一细菌属所主导。分类为呼吸道菌群的灌洗液与培养阳性样本在微生物组谱上更为相似。对组间细胞因子表达的比较表明,培养阳性和呼吸道菌群组中细胞因子(IFN-γ、IL-17F、IL-1β、IL-31、TNF-α)水平升高。培养阳性样本表现出更强的免疫反应和细菌属多样性降低。尽管培养阴性样本中的细胞因子水平较低,但细菌种类较多,这表明常驻非致病性细菌群落可能是正常肺部环境的标志。呼吸道菌群样本与培养阳性样本最为相似,在考虑临床治疗时,可能需要将其归类为培养阳性。