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Research Snapshot Theater: Infectious Disease, Pediatric I

Deciphering the Clinical Impact of Microbial Cell-Free DNA Sequencing in the Critically Ill Child

Tuesday, March 24, 2026
8:45 AM - 9:45 AM Central Time
Location: Connections Central - RST 05
  • IS

    Ian Santana, MD (he/him/his)

    Disclosure(s): No relevant financial relationship(s) to disclose.

  • AP

    Alejandro Paneque, MS2

    Medical Student
    UCF school of medicine, Nemours Children's Health Orlando

    Disclosure information not submitted.

  • LM

    Laura Mohr, CPNP

    Advanced Practice Provider
    Nemours Children's Hospital, Florida

    Disclosure information not submitted.

  • SV

    Sindy Villacres, DO, MEd

    Associate Designated Institutional Official, Pediatric critical care physician
    Nemours Childrens Hospital

    Disclosure information not submitted.

Introduction: Plasma microbial cell-free DNA sequencing tests, such as Karius, offer a noninvasive approach to microbial identification that may enable earlier diagnosis, targeted antimicrobial therapy, and reduced reliance on invasive procedures. However, current literature evaluating its clinical utility in critically ill pediatric populations remains limited, with existing studies constrained by small sample sizes.


Methods: A retrospective cohort study was conducted involving critically ill pediatric patients aged 0–18 years who were admitted to a freestanding tertiary care children’s hospital in Orlando, FL, between August 2019 and February 2025. Critical illness was defined by the presence of one or more of the following clinical criteria: (1) temperature instability, (2) hemodynamic alterations, (3) altered mental status, (4) abnormal perfusion, or (5) dysfunction of one or more organ systems. Detailed reviews were performed to compare results from standard infectious diagnostic tests (e.g., blood, urine, CSF, wound cultures), with those obtained from plasma microbial cell-free DNA sequencing (Karius). Provider documentation, including notes from infectious disease specialists and clinicians, were analyzed to assess the influence of Karius results on clinical management. Clinical impact was classified as a binary outcome, and statistical comparisons between standard testing and Karius results were conducted using Chi-square analysis with two-tailed p-values.


Results: A total of 199 Karius tests were reviewed. Of these, 60% (120/199) yielded positive results. Among the positive cases, 52% (62/120) demonstrated statistically significant clinical impact (p = 0.0001), resulting in changes to antimicrobial management. Within these 62 impactful cases, 47% (29/62) had negative standard microbial identification tests, while 53% (33/62) had positive standard results but Karius identified at least one additional organism not previously detected.


Conclusions: Plasma microbial cell-free DNA sequencing is a promising, noninvasive diagnostic tool that can detect pathogens missed by conventional methods. Its integration into clinical practice may enhance early identification of infections, guide antimicrobial therapy, and reduce the need for invasive diagnostics in critically ill pediatric populations.