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Updated: Mar 1, 2026

Novel Diagnostics in Revision Arthroplasty: Implant Sonication and Multiplex Polymerase Chain Reaction
Published on: December 3, 2017
Viable bacteria persist on antibiotic spacers following two-stage revision for periprosthetic joint infection
Dongzhu Ma1, Robert M Q Shanks2, Charles M Davis3
1Arthritis and Arthroplasty Design Group, Department of Orthopaedic Surgery, University of Pittsburgh, Pittsburgh, Pennsylvania, 15219.
Abstract:
Treatment in periprosthetic joint infection (PJI) remains challenging. The failure rate of two-stage revision and irrigation and debridement with component retention in PJI suggests that biofilm cells have a high tolerance to antibiotic chemotherapy. Previous work has demonstrated that biofilm cells have high antibiotic tolerance in vitro, but there is little clinical evidence to support these observations. The aim of this study was to determine if retrieved antibiotic spacers from two-stage revision total knee arthroplasty for PJI have evidence of remaining viable bacteria. Antibiotic poly (methyl methacrylate) (PMMA) spacers from two-stage revision total knee arthroplasty for PJI were prospectively collected and analyzed for bacterial 16s rRNA using polymerase chain reaction (PCR), reverse transcription (RT)-PCR, quantitative RT-PCR (qRT-PCR), and single genome analysis (SGA). PCR and RT-PCR identified bacterial species on 53.8% (7/13) of these samples. When initial culture negative cases are excluded, 68% (6/9) samples were identified with bacterial species. A more rigorous qRT-PCR analysis showed a strong positive signal for bacterial contamination in 30.7% (4/13) of cases. These patients did not show any clinical evidence of PJI recurrence after 15 months of follow-up. Because the half-life of bacterial rRNA is approximately a few days, the identification of bacteria rRNA on antibiotic PMMA spacers suggests that viable bacteria were present after conclusion of antibiotic therapy. This study provides evidence for the high tolerance of biofilm cells to antibiotics in vivo and the important role of bacterial persisters in PJI. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:452-458, 2018.
Insights
This study found viable bacteria on antibiotic spacers used in periprosthetic joint infection (PJI) treatment. This suggests bacteria in biofilms are highly tolerant to antibiotics, even in vivo.
Area of Science:
- Orthopaedic Surgery
- Infectious Diseases
- Biomaterials
Background:
- Periprosthetic joint infection (PJI) treatment is challenging due to high failure rates.
- Biofilm bacteria exhibit significant antibiotic tolerance in vitro, but clinical evidence is limited.
- Two-stage revision and irrigation/debridement have limitations in managing PJI.
Purpose of the Study:
- To investigate the presence of viable bacteria on antibiotic-eluting poly (methyl methacrylate) (PMMA) spacers after two-stage revision for PJI.
- To provide clinical evidence for antibiotic tolerance of biofilm bacteria in vivo.
- To explore the role of bacterial persisters in PJI treatment failure.
Main Methods:
- Prospective collection of antibiotic PMMA spacers from patients undergoing two-stage revision for PJI.
- Analysis of bacterial 16S rRNA using polymerase chain reaction (PCR), RT-PCR, and quantitative RT-PCR (qRT-PCR).
- Single genome analysis (SGA) was also employed for bacterial identification.
Main Results:
- Bacterial species were identified in 53.8% of samples by PCR/RT-PCR, increasing to 68% when initial culture-negative cases were excluded.
- Quantitative RT-PCR detected bacterial contamination in 30.7% of cases.
- No clinical evidence of PJI recurrence was observed in patients with positive bacterial rRNA findings after 15 months.
Conclusions:
- The presence of bacterial rRNA on antibiotic spacers suggests viable bacteria persist despite antibiotic therapy, indicating high in vivo antibiotic tolerance.
- Bacterial persister cells likely play a significant role in the challenges of treating PJI.
- Further research into strategies to eradicate persistent bacteria in PJI biofilms is warranted.
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