title: Use of 16S rRNA sequencing and quantitative PCR to correlate venous leg ulcer bacterial bioburden dynamics with wound expansion, antibiotic therapy, and healing.
abstract: 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.
authors.affiliation: Department of Microbiology and Immunology, Stanford University, Palo Alto, California.
authors.forename: Daniel D
authors.initials: DD
authors.lastname: Sprockett
authors.affiliation: Department of Dermatology, Case Western Reserve University, Cleveland, Ohio.
authors.forename: Christine G
authors.initials: CG
authors.lastname: Ammons
authors.affiliation: Department of Dermatology, Case Western Reserve University, Cleveland, Ohio.
authors.forename: Marie S
authors.initials: MS
authors.lastname: Tuttle
mesh_terms: D000328:Adult; D000900:Anti-Bacterial Agents / Q000627:therapeutic use*; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification; D058491:Bacterial Load; D004269:DNA, Bacterial / Q000235:genetics*; D003646:Debridement; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D009779:Occlusive Dressings; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D014647:Varicose Ulcer / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D014945:Wound Healing / Q000235:genetics*; D014946:Wound Infection / Q000188:drug therapy / Q000235:genetics* / Q000382:microbiology
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S