Abstract | March 11, 2024

A Comparative Analysis of Next-Generation Sequencing and Culture-Based Technology in Burn Microbiome Analysis

Henry Clayton Ross, BS, MS, 2nd Year Medical Student, Medical University of South Carolina, Charleston, SC

Caroline Corley, BS, MS3, MUSC, Charleston, SC; Carter M. Powell, BS, Mindy Engevik, Ph.D., Assistant Professor, Regenerative Medicine & Cell Biology, MUSC, Charleston, SC; Michael G. Schmidt, Ph.D., Professor, Microbiology & Immunology, MUSC, Charleston, SC; Lisa Steed, Ph.D., Professor, Pathology & Laboratory Medicine, MUSC, Charleston, SC; Arman Kilic, MD, Associate Professor, Surgery, MUSC, Charleston; Steven A. Kahn, MD, Associate Professor, Surgery, MUSC, Charleston, SC; Deepak K. Ozhathil, MD, Assistant Professor, Surgery, MUSC, Charleston, SC

Learning Objectives

  1. Identify the limitations of routine burn wound cultures and understand the potential of Next Generation Sequencing (NGS) as an alternative diagnostic method
  2. Assess the implications of NGS in burn care, recognizing its ability to capture diverse bacterial species and its potential to revolutionize treatment strategies based on microbial progression

Background/Knowledge Gap: Bacterial colonization of wounds is the primary contributor to graft loss and delayed wound healing in burn care. Multiple studies have linked bacterial counts with graft loss and specific microbes with delayed wound healing. Conventional microbial culture methods have yielded information on bacteria capable of colonizing wounds, but analysis of the complex microbial community in these wounds is lacking. Although Next Generation Sequencing (NGS) technology represents a promising alternative to traditional culturing, this technology has not been validated in burn wounds. Our study hypothesizes that NGS will be non-inferior to traditional culture-based methods in characterizing the wound microbiota.

Methods/Design: The study will enroll adults who undergo surgery for acute burn wounds. A pre-operative surface wound swab and two adjacent tissue specimens will be collected from 80 wounds. The swab and one tissue specimen will undergo 16S and 18S amplicon sequencing, while the other specimen will be sent for wound cultures. Species profiles and other outcomes will be compared with Chi-square and ANOVA/MANOVA tests using SPSS software.

Results/Findings: Preliminary results are available for 17 of 80 samples from 10 patients. Enrollees favor male gender, average 50.9 years of age, and suffered an average 16.6% total burned surface area injury. Samples were collected a mean of 15.1 days post-injury, cultures identified an average of one species and NGS identified three. Twelve specimens exhibited positive NGS results: 4 were culture negative, 6 captured all species detected on culture, and 2 had incomplete capture. Nine specimens identified species on NGS not detected on wound culture including some emerging pathogens. Of the 5 negative NGS specimens, 2 were positive by culture.

Conclusions/Implications: Our preliminary data support the predictive value of NGS relative to culture-based methods. Specimens collected closer to the date of injury were predominantly commensal/environmental species, while later specimens revealed enteric/pathogenic incursion. Pseudomonas aeruginosa predominance in tissue samples occurred after 14 days but was detected on swabs earlier, suggesting delayed tissue penetration. This study supports the use of NGS to characterize the burn wound microbiome and promotes its potential to uncover future biotherapeutic strategies.

References and Resources

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