Identification of prosthetic joint infections with 16S amplicon metagenomic sequencing - comparison with standard cultivation approach

Diagn Microbiol Infect Dis. 2024 May;109(1):116188. doi: 10.1016/j.diagmicrobio.2024.116188. Epub 2024 Jan 17.

Abstract

Prosthetic joint infections (PJIs) are commonly diagnosed via culture-based methods, which may miss hard-to-grow pathogens. This study contrasts amplicon metagenomic sequencing (16S AS) with traditional culture techniques for enhanced clinical decision-making. We analyzed sonicate fluid from 27 patients undergoing revision arthroplasty using both methods, emphasizing the distinction between contaminants and true positives. Our findings show moderate agreement between the two methods, with a Cohen's kappa of 0.490, varying across bacterial genera (Cohen's kappa -0.059 to 1). The sensitivity of 16S AS compared to culture was 81% (95% CI, 68% to 94%). Sequencing revealed greater microbial diversity, including anaerobic genera like Anaerococcus and Citrobacter. Interestingly, several culture-negative PJI samples showed diverse bacteria via 16S AS. Despite rigorous controls and algorithms to eliminate contaminants, confirming bacteria presence with 16S AS remains a challenge. This highlights the need for improved PJI diagnostic methods, while also pointing out the limitations of next-generation sequencing (NGS) as a clinical diagnostic tool.

Keywords: Amplicon metagenomic sequencing; Cultivation; Pathogen detection; Prosthetic joint infection.

MeSH terms

  • Arthritis, Infectious* / diagnosis
  • Arthroplasty
  • Bacteria / genetics
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Prostheses and Implants
  • Prosthesis-Related Infections* / diagnosis
  • Prosthesis-Related Infections* / microbiology
  • RNA, Ribosomal, 16S / genetics
  • Sensitivity and Specificity

Substances

  • RNA, Ribosomal, 16S