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Updated: Mar 11, 2026
In Situ Detection of Bacteria within Paraffin-embedded Tissues Using a Digoxin-labeled DNA Probe Targeting 16S rRNA
Published on: May 21, 2015
Neer Award 2017: A rapid method for detecting Propionibacterium acnes in surgical biopsy specimens from the shoulder
Scott Holmes1, Ana M Pena Diaz2, George S Athwal3
1Department of Medicine, Schulich School of Medicine and Dentistry, Western University, London, ON, Canada.
Background:
Propionibacterium (P) acnes infection of the shoulder after arthroplasty is a common and serious complication. Current detection methods for P acnes involve anaerobic cultures that require prolonged incubation periods (typically 7-14 days). We have developed a polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) approach that sensitively and specifically identifies P acnes in tissue specimens within a 24-hour period.
Methods:
Primers were designed to amplify a unique region of the 16S rRNA gene in P acnes that contained a unique HaeIII restriction enzyme site. PCR and RFLP analyses were optimized to detect P acnes DNA in in vitro cultures and in arthroscopic surgical biopsy specimens from patients with P acnes infections.
Results:
A 564 base-pair PCR amplicon was derived from all of the known P acnes strains. HaeIII digests of the amplicon yielded a restriction fragment pattern that was unique to P acnes. P acnes-specific amplicons were detected in as few as 10 bacterial cells and in clinical biopsy specimens of infected shoulder tissues.
Conclusion:
This PCR-RFLP assay combines the sensitivity of PCR with the specificity of RFLP mapping to identify P acnes in surgical isolates. The assay is robust and rapid, and a P acnes-positive tissue specimen can be confirmed within 24 hours of sampling, facilitating treatment decision making, targeted antibiotic therapy, and monitoring to minimize implant failure and revision surgery.
Insights
A new polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay rapidly and accurately detects Propionibacterium acnes infections in shoulder arthroplasty patients. This method significantly shortens diagnostic time from days to 24 hours.
Area of Science:
- Microbiology
- Molecular Diagnostics
- Orthopedic Surgery
Background:
- Propionibacterium acnes (P. acnes) infections are a significant complication following shoulder arthroplasty.
- Current diagnostic methods rely on anaerobic cultures with lengthy incubation periods (7-14 days).
Purpose of the Study:
- To develop a rapid and specific diagnostic assay for P. acnes in surgical specimens.
- To reduce the time to diagnosis for P. acnes infections post-arthroplasty.
Main Methods:
- Development of a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay targeting the 16S rRNA gene of P. acnes.
- Optimization of PCR and RFLP for detecting P. acnes DNA in vitro and in clinical biopsy samples.
Main Results:
- A unique 564 base-pair PCR amplicon specific to P. acnes was identified.
- HaeIII restriction enzyme digestion produced a pattern unique to P. acnes.
- The assay detected P. acnes in as few as 10 bacterial cells and in infected clinical specimens.
Conclusions:
- The PCR-RFLP assay offers a sensitive and specific method for identifying P. acnes in surgical isolates.
- This rapid assay provides results within 24 hours, aiding timely treatment decisions and potentially minimizing implant failure and revision surgery.
