Amplification-Based DNA Analysis in the Diagnosis of Prosthetic Joint Infection

Abstract

Microbiological cultures are moderately sensitive for diagnosing prosthetic joint infection (PJI). This study was conducted to determine whether amplificationbased DNA methods applied on intraoperative samples could enhance PJI diagnosis compared with culture alone in routine surgical practice. Revision arthroplasty was performed for suspected PJI (n ‫؍‬ 41) and osteoarthrosis control (n ‫؍‬ 28) patients , and a diagnosis of PJI was confirmed in 34 patients. Amplification by polymerase chain reaction was performed on both 16S ribosomal DNA universal target genes and femA Staphylococcus-specific target genes. Species identification was achieved through amplicon sequencing. Amplification of the femA gene led to subsequent testing for methicillin resistance by amplification of the mecA gene. Microbiological and molecular assays identified a causative organism in 22 of 34 patients (64.7%) and in 31 of 34 patients (91.2%), respectively. In 18 of the 22 culture-positive patients, molecular and microbiological results were concordant for bacterial genus , species , and/or methicillin resistance. Bacterial agents were identified only by molecular methods in nine PJI patients , including seven who were receiving antibiotics at the time of surgery and one with recent but not concomitant antibiotherapy. DNA-based methods were found to effectively complement microbiological methods , without interfering with existing procedures for sample collection , for the identification of causative pathogens from intraoperative PJI samples , especially in patients with recent or concomitant antibiotherapy. (J Mol

Diagnosis of prosthetic joint infections using UMD-Universal Kit and the automated multiplex-PCR Unyvero i60 ITI® cartridge system: a pilot study

“…16S rRNA gene real-time PCR results were in line with the conventional culture results in about 96 % of the studied patient cases. These findings seem to be consistent with previously published data on NAAT techniques, indicating a relatively wide range of sensitivity and specificity [8][9][10][11][12]. NAATs may reasonably expand the diagnostic panel and contribute to a higher recovery rate especially after antimicrobial use.…”

“…Only few studies have assessed the diagnostic utility of nucleic acid amplification test (NAAT) techniques in this context. The reported findings for different PJI-associated materials indicate a wide range of sensitivity (50-92 %) and specificity (65-94 %) [8][9][10][11][12][13]. Results from a recent study comparing 16S rRNA gene PCR and consecutive sequencing of sonication fluid with culture of synovial aspirates, tissue samples and sonication fluids reports that 16S rRNA gene PCR improved sensitivity [14].…”

First Experience of a Multicenter External Quality Assessment of Molecular 16S rRNA Gene Detection in Bone and Joint Infections

“…Molecular biology provided a diagnostic tool that showed its effectiveness, especially when antibiotics were administered before surgery or when fastidious bacteria were involved, particularly for the diagnosis of endocarditis (4). For BJI diagnosis, broad-range 16S rRNA gene PCR has shown higher, lower, or equivalent sensitivity compared with conventional culture methods, but sometimes to the detriment of specificity (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15).…”

Infection in Primary Hip and Knee Arthroplasty

“…In a review of 34 patients with confirmed prosthetic joint infection Vandercam et al found that PCR was positive in 31 of 34 patients (91.2%), compared to positive microbiological culture in 22 of 34 patients (64.7%). Of import, eight of the nine patients with positive PCR but negative culture results had received antibiotic therapy in the prior ten days (Vandercam, et al 2008). Despite these promising results, the weakness of 16s ribosomal RNA PCR techniques is the low specificity and high false positive rate.…”