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Novel Diagnostics in Revision Arthroplasty: Implant Sonication and Multiplex Polymerase Chain Reaction
Published on: December 3, 2017
Amplification-based DNA analysis in the diagnosis of prosthetic joint infection
Bernard Vandercam1, Sabine Jeumont, Olivier Cornu
1Division of Infectious Diseases, St-Luc's University Hospital, Université catholique de Louvain, Brussels, Belgium.
Abstract:
Microbiological cultures are moderately sensitive for diagnosing prosthetic joint infection (PJI). This study was conducted to determine whether amplification-based 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.
Insights
DNA-based molecular methods significantly improve prosthetic joint infection (PJI) diagnosis compared to traditional cultures. These amplification-based techniques enhance pathogen identification, particularly in patients undergoing revision arthroplasty with prior antibiotic exposure.
Area of Science:
- Orthopedic Surgery
- Infectious Diseases
- Molecular Diagnostics
Background:
- Prosthetic joint infection (PJI) diagnosis relies on microbiological cultures, which have moderate sensitivity.
- Enhancing diagnostic accuracy for PJI is crucial for effective treatment and patient outcomes.
Purpose of the Study:
- To evaluate if amplification-based DNA methods on intraoperative samples improve PJI diagnosis compared to culture alone.
- To assess the utility of molecular techniques in routine surgical practice for PJI detection.
Main Methods:
- Revision arthroplasty patients (n=41) with suspected PJI and osteoarthrosis controls (n=28) were included.
- Polymerase chain reaction (PCR) amplified 16S ribosomal DNA and Staphylococcus-specific femA genes.
- Amplicon sequencing identified species, and mecA gene amplification detected methicillin resistance.
Main Results:
- Molecular assays identified causative organisms in 91.2% (31/34) of PJI patients, versus 64.7% (22/34) for cultures.
- Molecular methods alone identified pathogens in nine PJI patients, including those with recent or ongoing antibiotic treatment.
- Concordance between molecular and microbiological results was high for bacterial identification and antibiotic resistance.
Conclusions:
- DNA-based molecular methods effectively complement microbiological cultures for identifying pathogens in intraoperative PJI samples.
- These molecular techniques are particularly valuable in patients with recent or concomitant antibiotherapy, improving PJI diagnosis.
- Molecular diagnostics can be integrated into existing sample collection procedures without interference.
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