Plasma Microbial Cell-free DNA Sequencing for the Detection of Kingella kingae Pediatric Spinal Infections

Pediatr Infect Dis J. 2025 Jun 1;44(6):582-588. doi: 10.1097/INF.0000000000004743. Epub 2025 Feb 21.

Abstract

Background: Diagnosis of Kingella kingae skeletal system infections is made challenging by the microbe's fastidious nature. Detection and quantification of circulating microbial cell-free DNA (mcfDNA) in plasma by the Karius Test, a commercial metagenomic sequencing test, may offer promise in diagnosing pediatric spinal infections caused by difficult-to-culture organisms such as K. kingae .

Methods: Plasma mcfDNA sequencing detections of K. kingae from April 2018 to December 2020 were reviewed to identify pediatric (age <18 years) patients. Medical charts of those with spinal infections were reviewed, and mcfDNA sequencing diagnostic performance was compared with usual care tests (ie, cultures, polymerase chain reaction).

Results: Ten children with K. kingae spinal infections were identified across 7 institutions. The median age was 16.5 months (range 11-23 months). All case-patients had vertebral osteomyelitis with 9 having spondylodiscitis. Compared with usual care tests, mcfDNA sequencing was significantly more sensitive (McNemar's test 6.25, 2-tailed P = 0.0133). It was the only method of microbiological diagnosis in 9 patients, providing results in a median of 2.5 days (range 2-5 days) from sample collection. K. kingae mcfDNA was detected despite antibiotic pretreatment in 5/5 case-patients. Pathogen-tailoring of antimicrobial coverage was undertaken in 9 children.

Conclusion: Plasma mcfDNA sequencing offers a rapid, noninvasive method of detecting K. kingae causing pediatric spinal infections. This culture-independent approach may facilitate diagnosis, despite antibiotic pretreatment and subsequently targeted therapy and potentially obviate the need for biopsy.

Keywords: diagnostics; metagenomic sequencing; pediatrics; spinal infection.

MeSH terms

  • Cell-Free Nucleic Acids* / blood
  • Cell-Free Nucleic Acids* / genetics
  • Child
  • Child, Preschool
  • DNA, Bacterial* / blood
  • DNA, Bacterial* / genetics
  • Discitis* / diagnosis
  • Discitis* / microbiology
  • Female
  • Humans
  • Infant
  • Kingella kingae* / genetics
  • Kingella kingae* / isolation & purification
  • Male
  • Neisseriaceae Infections* / blood
  • Neisseriaceae Infections* / diagnosis
  • Neisseriaceae Infections* / microbiology
  • Osteomyelitis* / diagnosis
  • Osteomyelitis* / microbiology
  • Retrospective Studies
  • Sensitivity and Specificity
  • Sequence Analysis, DNA / methods

Substances

  • DNA, Bacterial
  • Cell-Free Nucleic Acids