PROOF OF CONCEPT OF SHOTGUN-METAGENOMICS ON BLOOD CULTURE BOTTLES INOCULATED WITH PROSTHETIC JOINT TISSUE. SHOTGUN-METAGENOMICS ON BLOOD CULTURE BOTTLES WITH TISSUE

Clinical metagenomics is actively moving from research to clinical laboratories. It has the potential to change the microbial diagnosis of infectious diseases, especially when the detection and identification of pathogens can be challenging, such as prosthetic joint infection (PJI). The application of shotgun-metagenomics (SMg) to periprosthetic joint tissue (PJT) specimens is often challenged by low loads of bacteria in addition to high inhibitor and contaminant host DNA limiting pathogen recovery. SMg performed directly from blood culture bottles (BCBs) inoculated with PJT may be a convenient approach for addressing these obstacles. Our aim is to evaluate a method for the preparation of DNA suitable for SMg directly from BCBs inoculated with PJT specimens and assess their performances through metagenomic sequencing for the identification of pathogens. We test shotgun-metagenomics on BCB inoculated with: PJT clinical samples from patients with suspicion of PJI, samples spiked with bacterial species related with PJI and one sample inoculated with sterilized tissue (negative control) to get a comprehensive overview.