Sprockett Daniel D, Ammons Christine G, Tuttle Marie S
Department of Microbiology and Immunology, Stanford University, Palo Alto, California.
Department of Dermatology, Case Western Reserve University, Cleveland, Ohio.
Wound Repair Regen. 2015 Sep;23(5):765-71. doi: 10.1111/wrr.12309. Epub 2015 Jun 24.
Clinical diagnosis of infection in chronic wounds is currently limited to subjective clinical signs and culture-based methods that underestimate the complexity of wound microbial bioburden as revealed by DNA-based microbial identification methods. Here, we use 16S rRNA next generation sequencing and quantitative polymerase chain reaction to characterize weekly changes in bacterial load, community structure, and diversity associated with a chronic venous leg ulcer over the 15-week course of treatment and healing. Our DNA-based methods and detailed sampling scheme reveal that the bacterial bioburden of the wound is unexpectedly dynamic, including changes in the bacterial load and community structure that correlate with wound expansion, antibiotic therapy, and healing. We demonstrate that these multidimensional changes in bacterial bioburden can be summarized using swabs taken prior to debridement, and therefore, can be more easily collected serially than debridement or biopsy samples. Overall, this case illustrates the importance of detailed clinical indicators and longitudinal sampling to determine the pathogenic significance of chronic wound microbial dynamics and guide best use of antimicrobials for improvement of healing outcomes.
慢性伤口感染的临床诊断目前仅限于主观临床体征和基于培养的方法，这些方法低估了基于DNA的微生物鉴定方法所揭示的伤口微生物生物负荷的复杂性。在此，我们使用16S rRNA下一代测序和定量聚合酶链反应来表征在15周的治疗和愈合过程中，与慢性下肢静脉溃疡相关的细菌载量、群落结构和多样性的每周变化。我们基于DNA的方法和详细的采样方案表明，伤口的细菌生物负荷出人意料地具有动态性，包括与伤口扩大、抗生素治疗和愈合相关的细菌载量和群落结构变化。我们证明，这些细菌生物负荷的多维变化可以用清创术前采集的拭子进行总结，因此，与清创术或活检样本相比，更容易连续采集。总体而言，本病例说明了详细的临床指标和纵向采样对于确定慢性伤口微生物动态的致病意义以及指导抗菌药物的最佳使用以改善愈合结果的重要性。
