Division of Infectious Diseases, Geneva University Hospitals and Faculty of Medicine, University of Geneva, 4 Rue Gabrielle-Perret-Gentil, 1211, Geneva, Switzerland.
Bacteriology Laboratory, Geneva University Hospitals and Faculty of Medicine, University of Geneva, Geneva, Switzerland.
Intensive Care Med. 2019 Aug;45(8):1082-1092. doi: 10.1007/s00134-019-05660-8. Epub 2019 Jun 17.
To compare bacteria recovered by standard cultures and metataxonomics, particularly with regard to ventilator-associated pneumonia (VAP) pathogens, and to determine if the presence of particular bacteria or microbiota in tracheal and oropharyngeal secretions during the course of intubation was associated with the development of VAP.
In this case-control study, oropharyngeal secretions and endotracheal aspirate were collected daily in mechanically ventilated patients. Culture and metataxonomics (16S rRNA gene-based taxonomic profiling of bacterial communities) were performed on serial upper respiratory samples from patients with late-onset definite VAP and their respective controls.
Metataxonomic analyses showed that a low relative abundance of Bacilli at the time of intubation in the oropharyngeal secretions was strongly associated with the subsequent development of VAP. On the day of VAP, the quantity of human and bacterial DNA in both tracheal and oropharyngeal secretions was significantly higher in patients with VAP than in matched controls with similar ventilation times. Molecular techniques identified the pathogen(s) of VAP found by culture, but also many more bacteria, classically difficult to culture, such as Mycoplasma spp. and anaerobes.
Molecular analyses of respiratory specimens identified markers associated with the development of VAP, as well as important differences in the taxa abundance between VAP and controls. Further prospective trials are needed to test the predictive value of these markers, as well as the relevance of uncultured bacteria in the pathogenesis of VAP.
比较标准培养物和宏基因组学方法所分离的细菌,尤其是呼吸机相关性肺炎(VAP)病原体,同时明确气管和口咽分泌物中特定细菌或微生物群落的存在与 VAP 发展之间的关系。
本病例对照研究对机械通气患者的口咽分泌物和气管内吸出物进行了每日采集。对发生迟发性明确 VAP 的患者及其各自对照者的上呼吸道连续样本进行培养和宏基因组学(基于 16S rRNA 基因的细菌群落分类分析)检测。
宏基因组分析表明,口咽分泌物中 Bacilli 的相对丰度较低与随后发生 VAP 有很强的相关性。发生 VAP 当天,VAP 患者的气管和口咽分泌物中人体和细菌 DNA 的量均显著高于通气时间相似的匹配对照者。分子技术不仅鉴定了培养法发现的 VAP 病原体,而且还鉴定了许多更难培养的细菌,如支原体属和厌氧菌。
对呼吸道标本进行的分子分析确定了与 VAP 发展相关的标志物,以及 VAP 和对照组之间分类群丰度的重要差异。需要进一步前瞻性试验来检验这些标志物的预测价值,以及未培养细菌在 VAP 发病机制中的相关性。