Differential Contributions of Specimen Types, Culturing, and 16S rRNA Sequencing in Diagnosis of Prosthetic Joint Infections

Abstract

Prosthetic joint failure is mainly caused by infection, aseptic failure (AF), and mechanical problems. Infection detection has been improved with modified culture methods and molecular diagnostics. However, comparisons between modified and conventional microbiology methods are difficult due to variations in specimen sampling. In this prospective, multidisciplinary study of hip or knee prosthetic failures, we assessed the contributions of different specimen types, extended culture incubations, and 16S rRNA sequencing for diagnosing prosthetic joint infections (PJI). Project specimens included joint fluid (JF), bone biopsy specimens (BB), soft-tissue biopsy specimens (STB), and swabs (SW) from the prosthesis, collected in situ, and sonication fluid collected from prosthetic components (PC). Specimens were cultured for 6 (conventional) or 14 days, and 16S rRNA sequencing was performed at study completion. Of the 156 patients enrolled, 111 underwent 114 surgical revisions (cases) due to indications of either PJI (n = 43) or AF (n = 71). Conventional tissue biopsy cultures confirmed PJI in 28/43 (65%) cases and refuted AF in 3/71 (4%) cases; one case was not evaluable. Based on these results, minor diagnostic adjustments were made. Fourteen-day cultures of JF, STB, and PC specimens confirmed PJI in 39/42 (93%) cases, and 16S rRNA sequencing confirmed PJI in 33/42 (83%) cases. One PJI case was confirmed with 16S rRNA sequencing alone and five with cultures of project specimens alone. These findings indicated that JF, STB, and PC specimen cultures qualified as an optimal diagnostic set. The contribution of sequencing to diagnosis of PJI may depend on patient selection; this hypothesis requires further investigation.

Multidisciplinary Diagnostic Algorithm for Evaluation of Patients Presenting with a Prosthetic Problem in the Hip or Knee: A Prospective Study

“…We foresee that results obtained with 16S rRNA gene PCR followed by amplicon sequencing may help to clarify this latter group. Data from culture and 16S rRNA in the different specimen types are published elsewhere [36].…”

72 revision surgeries for aseptic failure after hip or knee arthroplasty: a prospective study with an extended diagnostic algorithm

“…In the present prospective study, we report a cohort of patients undergoing revision and investigated for AF according to a diagnostic algorithm and undergoing revision surgery [23]. We evaluated results for 71 patients undergoing revision surgery for AF after THA and TKA within a larger study addressing the diagnosis of prosthesis-related infection and pain.…”

“…Results from the different studies including this performed under the PRIS project did not support a significant value of routine molecular tests of intraoperative tissue in patients suspected of AF. Details from 16S rRNA in this study have been published elsewhere [23].…”

“…Patients were invited to participate in a study with the primary aim to improve the diagnosis of prosthesis-related infection and pain (Danish acronym PRIS) through a multidisciplinary diagnostic algorithm [23]. The project was carried out between 2011 and 2014 in the North Denmark Region, with a population of approximately 580.000.…”

“…To support these elaborate procedures all containers and transport media were provided as a set in a box [25]. Transport and handling of specimens including sonication of prosthetic components have been described elsewhere [23, 25]. Triplicates made it feasible to perform parallel testing by bacteriological culturing for 14 days (routine 6 days) and 16S rRNA gene amplification followed by amplicon sequencing (22).…”