Molecular analysis of oral and respiratory bacterial species associated with ventilator-associated pneumonia
Molecular analysis of oral and respiratory bacterial species associated with ventilator-associated pneumonia
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DOI：
10.1128/jcm.01963-06
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发表时间：
2007-05-01
影响因子：
9.4
通讯作者：
Lockhart, Peter B.
中科院分区：
文献类型：
--
作者：
Bahrani-Mougeot, Farah K.;Paster, Bruce J.;Lockhart, Peter B.
需要机械呼吸支持的创伤重症监护病房（TICU）患者经常发生呼吸机相关性肺炎（VAP）。口腔和口咽细菌被认为是许多VAP病例的原因，但缺乏这种关系的确切证据。早期的研究使用传统的基于培养的方法来鉴定细菌病原体，但这些方法是不够的，因为一些细菌可能是不可培养的或难以生长。本研究的目的是使用一种不依赖于培养的分子方法来分析和比较TICU患者发生VAP后口腔和肺部的细菌种类。16例患者的背舌和支气管肺泡灌洗液中获得细菌样本。提取细菌DNA，PCR扩增16 S rRNA基因，克隆到大肠杆菌中，并测序。测序数据揭示了以下内容：（i）口腔和肺部部位的细菌种类的广泛多样性，其中一些是新的;（ii）已知和推定的呼吸道病原体定殖于14名患者的口腔和肺部;以及（iii）许多细菌病原体（例如，肺部样本中存在肺炎双球菌、缓慢嗜血杆菌、麻疹双球菌和荧光假单胞菌），该中心之前使用基于培养的方法时尚未报告过这些情况。我们的数据表明，舌背表面作为一个潜在的水库细菌物种参与VAP。此外，很明显，VAP的细菌病原体的多样性远比目前的文献所表明的要复杂得多。
Trauma intensive care unit (TICU) patients requiring mechanical respiratory support frequently develop ventilator-associated pneumonia (VAP). Oral and oropharyngeal bacteria are believed to be responsible for many cases of VAP, but definitive evidence of this relationship is lacking. Earlier studies used conventional culture-based methods for identification of bacterial pathogens, but these methods are insufficient, as some bacteria may be uncultivable or difficult to grow. The purpose of this study was to use a culture-independent molecular approach to analyze and compare the bacterial species colonizing the oral cavity and the lungs of TICU patients who developed VAP. Bacterial samples were acquired from the dorsal tongue and bronchoalveolar lavage fluid of 16 patients. Bacterial DNA was extracted, and the 16S rRNA genes were PCR amplified, cloned into Escherichia coli, and sequenced. The sequencing data revealed the following: (i) a wide diversity of bacterial species in both the oral and pulmonary sites, some of them novel; (ii) known and putative respiratory pathogens colonizing both the oral cavity and lungs of 14 patients; and (iii) a number of bacterial pathogens (e.g., Dialister pneumosintes, Haemophilus segnis, Gemella morbillorum, and Pseudomonas fluorescens) in lung samples that had not been reported previously at this site when culture-based methods were used. Our data indicate that the dorsal surface of the tongue serves as a potential reservoir for bacterial species involved in VAP. Furthermore, it is clear that the diversity of bacterial pathogens for VAP is far more complex than the current literature suggests.