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Updated: May 3, 2026
Novel Diagnostics in Revision Arthroplasty: Implant Sonication and Multiplex Polymerase Chain Reaction
Published on: December 3, 2017
Molecular diagnosis of periprosthetic joint infection by quantitative RT-PCR of bacterial 16S ribosomal RNA
Mel S Lee1, Wen-Hsin Chang2, Su-Chin Chen3
1Department of Orthopaedic Surgery, Chia-Yi Chang Gung Memorial Hospital, 6 West Sec., Chiapu Road, Putzu City, Chiayi County 613, Taiwan ; Department of Orthopaedic Surgery, Linkou Chang Gung Memorial Hospital, Putzu City, Chiayi County 613, Taiwan ; College of Medicine, Chang Gung University, Putzu City, Chiayi County 613, Taiwan.
Abstract:
The diagnosis of periprosthetic joint infection is sometimes straightforward with purulent discharge from the fistula tract communicating to the joint prosthesis. However it is often difficult to differentiate septic from aseptic loosening of prosthesis because of the high culture-negative rates in conventional microbiologic culture. This study used quantitative reverse transcription polymerase chain reaction (RT-qPCR) to amplify bacterial 16S ribosomal RNA in vitro and in 11 clinical samples. The in vitro analysis demonstrated that the RT-qPCR method was highly sensitive with the detection limit of bacterial 16S rRNA being 0.148 pg/ μ l. Clinical specimens were analyzed using the same protocol. The RT-qPCR was positive for bacterial detection in 8 culture-positive cases (including aerobic, anaerobic, and mycobacteria) and 2 culture-negative cases. It was negative in one case that the final diagnosis was confirmed without infection. The molecular diagnosis of bacterial infection using RT-qPCR to detect bacterial 16S rRNA around a prosthesis correlated well with the clinical findings. Based on the promising clinical results, we were attempting to differentiate bacterial species or drug-resistant strains by using species-specific primers and to detect the persistence of bacteria during the interim period before the second stage reimplantation in a larger scale of clinical subjects.
Insights
Quantitative reverse transcription polymerase chain reaction (RT-qPCR) accurately detects bacterial 16S ribosomal RNA for diagnosing periprosthetic joint infections, even in culture-negative cases. This molecular method shows promise for improved infection diagnosis.
Area of Science:
- Microbiology
- Molecular Biology
- Orthopedic Surgery
Background:
- Diagnosing periprosthetic joint infection (PJI) is challenging due to high rates of culture-negative results with conventional methods.
- Differentiating septic from aseptic prosthesis loosening is critical for effective patient management.
Purpose of the Study:
- To evaluate the efficacy of quantitative reverse transcription polymerase chain reaction (RT-qPCR) for detecting bacterial 16S ribosomal RNA in periprosthetic joint infections.
- To assess the correlation of RT-qPCR results with clinical findings in suspected PJI cases.
Main Methods:
- Bacterial 16S ribosomal RNA was amplified using RT-qPCR in vitro and in 11 clinical periprosthetic samples.
- The sensitivity of the RT-qPCR method was determined through in vitro analysis.
- Clinical specimens were analyzed using the established RT-qPCR protocol.
Main Results:
- The RT-qPCR assay demonstrated high sensitivity with a detection limit of 0.148 pg/μl for bacterial 16S rRNA.
- RT-qPCR correctly identified bacterial infection in 8 culture-positive and 2 culture-negative cases.
- The method yielded a negative result in a non-infected control case, aligning with clinical diagnosis.
Conclusions:
- RT-qPCR detection of bacterial 16S rRNA is a sensitive and reliable molecular diagnostic tool for periprosthetic joint infections.
- This molecular approach correlates well with clinical findings and offers potential for improved diagnosis in challenging cases.
- Future research aims to expand RT-qPCR for bacterial species identification and monitoring treatment efficacy.
