Benefits of Polymerase Chain Reaction Combined With Culture for the Diagnosis of Bone and Joint Infections: A Prospective Test Performance Study

Abstract

BackgroundThe microbiological diagnosis of bone and joint infections (BJI) currently relies on cultures, and the relevance of molecular methods is still debated. The aim of this study was to determine whether polymerase chain reaction (PCR) could improve the etiological diagnosis of BJI.MethodsA prospective study was conducted during a 4-year period at Lariboisiere University Hospital (Paris, France), including patients with suspicion of infectious spondylodiscitis, septic arthritis, prosthetic joint infections, and respective noninfected groups. Clinical and radiological data were collected at inclusion and during follow-up. All samples were analyzed by conventional cultures and 16S ribosomal deoxyribonucleic acid (rDNA) gene (16S-PCR). Specific cultures and PCR targeting Mycobacterium tuberculosis were also performed for spondylodiscitis samples. Case records were subsequently analyzed by an independent expert committee to confirm or invalidate the suspicion of infection and definitively classify the patients in a case or control group. The sensitivity of the combination of culture and PCR was compared with culture alone.ResultsAfter expert committee analysis, 105 cases of BJI cases and 111 control patients were analyzed. The most common pathogens of BJI were staphylococci (30%), M tuberculosis (19%), and streptococci (14%). Adding PCR enhanced the sensitivity compared with culture alone (1) for the diagnosis of M tuberculosis spondylodiscitis (64.4% vs 42.2%; P < .01) and (2) for nonstaphylococci BJI (81.6% vs 71.3%; P < .01). It is interesting to note that 16S-PCR could detect BJI due to uncommon bacteria such as Mycoplasma and fastidious bacteria.ConclusionsOur study showed the benefit of 16S-PCR and PCR targeting M tuberculosis as add-on tests in cases of suspected BJI.

Evaluation of a syndromic panel polymerase chain reaction (spPCR) assay for the diagnosis of device‐associated bone and joint infections (BJI)

“…By providing increased accuracy in pathogen detection, results of amplification-based, molecular assays may support this decision-making process by providing critical information on causative pathogens in addition to conventional culture-based techniques. In fact, a previous study has reported a higher sensitivity of culture in combination with PCR than that of culture alone to detect infection-causing organisms (95.5% vs 81.8%, (Jacquier et al, 2019). In line with previous reports (Bemer et al, 2014) our study provides evidence that 16S rDNA PCR approaches to culture-independent pathogen detection is of apparent low sensitivity, with an estimated sensitivity of 37.5% compared to SOC.…”

“…Use of a spPCR showed increased pathogen yield compared to a 16S rDNA PCR, and this was also true for the detection of staphylococci, having found to be specifically challenging for 16S rDNA PCR assays (Jacquier et al, 2019). Thus, spPCR might prove as a reasonable addition to culture-based pathogen detection strategies in BJI.…”

Sonication versus the conventional method for evaluation of the dental microbiome: a prospective pilot study

“…Focusing on the study design, neither orthopaedic nor else studies, which are comparing direct 16S-rRNAgene analysis and the difference in the microbiome distribution in different sampling modalities exist. Only a few studies have been investigating, whether using 16S-rRNA-gene analysis resulted in similar or even improved results, than normal microbiological testing [26,[44][45][46]. Due to this fact and the appropriate results of 16S-rRNA-gene analysis in microbiome analysis of the oral cavity, we assume, that this is a reliable tool for such investigation [30,32,34,47].…”

Sampling challenges in diagnosis of chronic bacterial infections

“…These include (i) the fact that they cannot be used to measure antimicrobial susceptibility, (ii) the introduction of bias through the use of general primers for 16 s rRNA sequencing that do not bind equally to all bacteria and (iii) the difficulty in identification beyond the genus due to high similarity among closely related species. Several studies have compared the use of culturing and amplicon sequencing for diagnostics, generally reporting a higher sensitivity for sequencing compared to culturing, especially in the investigation of chronic infections [62][63][64]. If sequencing is to replace culturing as a clinical diagnostic method, it must be standardized, automated and cost-efficient.…”