The importance of a multifaceted approach to characterizing the microbial flora of chronic wounds
The importance of a multifaceted approach to characterizing the microbial flora of chronic wounds
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DOI：
10.1111/j.1524-475x.2011.00720.x
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发表时间：
2011-09-01
影响因子：
2.9
通讯作者：
Lazarus, Gerald S.
中科院分区：
文献类型：
--
作者：
Han, Anne;Zenilman, Jonathan M.;Lazarus, Gerald S.
慢性伤口含有复杂的固着生物体的多微生物群落，由于标准培养技术的限制，这些生物体一直未得到充分的重视。这项工作的目的是联合收割机最近开发的下一代调查技术，全面描述慢性伤口的微生物特征。组织样本来自约翰霍普金斯伤口中心的15名慢性伤口患者。标准细菌学培养显示伤口样本中平均有三种常见细菌。相比之下，高通量焦磷酸测序显示细菌多样性增加，每个伤口中平均有17个属。将慢性伤口的微生物群落分析数据与已发表的正常皮肤细菌测序分析进行比较。在慢性伤口中发现厌氧菌、革兰氏阴性杆菌和革兰氏阳性球菌的比例增加。此外，与正常皮肤相比，慢性伤口具有显著较低的丙酸杆菌种群。使用落射荧光显微镜，伤口细菌在高度组织化的厚汇合生物膜中或分散的单个细菌细胞中可视化。荧光原位杂交允许伤口样本中的金黄色葡萄球菌细胞的可视化。通过生物测定法测量群体感应分子以评估伤口中细菌之间的信号模式。在伤口样本中检测到一系列自诱导物-2活性。总的来说，这些数据为慢性伤口中细菌的身份，组织和行为提供了新的见解。这些信息可能为未来伤口愈合的有效策略提供重要线索。
Chronic wounds contain complex polymicrobial communities of sessile organisms that have been underappreciated because of limitations of standard culture techniques. The aim of this work was to combine recently developed next-generation investigative techniques to comprehensively describe the microbial characteristics of chronic wounds. Tissue samples were obtained from 15 patients with chronic wounds presenting to the Johns Hopkins Wound Center. Standard bacteriological cultures demonstrated an average of three common bacterial species in wound samples. By contrast, high-throughput pyrosequencing revealed increased bacterial diversity with an average of 17 genera in each wound. Data from microbial community profiling of chronic wounds were compared with published sequenced analyses of bacteria from normal skin. Increased proportions of anaerobes, Gram-negative rods and Gram-positive cocci were found in chronic wounds. In addition, chronic wounds had significantly lower populations of Propionibacterium compared with normal skin. Using epifluorescence microscopy, wound bacteria were visualized in highly organized thick confluent biofilms or as scattered individual bacterial cells. Fluorescent in situ hybridization allowed for the visualization of Staphylococcus aureus cells in a wound sample. Quorum-sensing molecules were measured by bioassay to evaluate signaling patterns among bacteria in the wounds. A range of autoinducer-2 activities was detected in the wound samples. Collectively, these data provide new insights into the identity, organization, and behavior of bacteria in chronic wounds. Such information may provide important clues to effective future strategies in wound healing.