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Novel Diagnostics in Revision Arthroplasty: Implant Sonication and Multiplex Polymerase Chain Reaction
Published on: December 3, 2017
Comparison of molecular and culture method in diagnosis of prosthetic joint infection
Mitja Rak1, Darja Barlič-Maganja, Martina Kavčič
1Faculty of Health Sciences, University of Primorska, Izola, Slovenia. mitja.rak@fvz.upr.si
Abstract:
Diagnosis of prosthetic joint infection with culture technique can be problematic since the causative agent(s) are not possible to cultivate in all cases. Molecular methods had been evaluated in many studies but their inclusion in routine diagnostics is still controversial. The purpose of our prospective study was to compare the diagnostic accuracy of broad-range (BR)-PCR and culture technique. Intraoperative samples of periprosthetic tissue were retrieved in 67 patients undergoing revision arthroplasty. Samples were analyzed with culture technique, immunohistochemistry and BR 16S rRNA gene PCR. Bacteria in PCR-positive samples were identified using two different methods: direct sequencing of PCR products and specific TaqMan assays. In 63 cases, full concordance was found between BR-PCR and culture technique. Specific TaqMan assays failed to identify bacteria in four culture- and BR-PCR-positive cases and therefore had a lower sensitivity in comparison with BR-PCR. Molecular methods detected bacteria with the same accuracy as culture; however, identification of bacteria was inferior to culture. Further development of species-recognition techniques is required to improve identification of causative microorganisms.
Insights
Broad-range PCR (BR-PCR) shows similar accuracy to culture for diagnosing prosthetic joint infections. However, culture remains superior for identifying specific bacteria, highlighting the need for improved molecular identification techniques.
Area of Science:
- Microbiology
- Orthopedic Surgery
- Molecular Diagnostics
Background:
- Diagnosing prosthetic joint infection (PJI) using traditional culture techniques is often limited by the inability to cultivate causative microorganisms in all cases.
- The clinical utility of molecular diagnostic methods for PJI remains a subject of ongoing debate and requires further investigation for routine implementation.
Purpose of the Study:
- To prospectively compare the diagnostic accuracy of broad-range PCR (BR-PCR) against conventional culture techniques for identifying pathogens in patients with PJI.
- To evaluate the effectiveness of direct sequencing and TaqMan assays for bacterial identification in BR-PCR positive samples.
Main Methods:
- Intraoperative periprosthetic tissue samples were collected from 67 patients undergoing revision arthroplasty.
- Samples were analyzed using standard culture methods, immunohistochemistry, and broad-range 16S rRNA gene PCR.
- Bacterial identification in PCR-positive samples was performed via direct sequencing and specific TaqMan assays.
Main Results:
- A high degree of concordance (63 out of 67 cases) was observed between BR-PCR and culture techniques in detecting bacterial presence.
- Specific TaqMan assays demonstrated lower sensitivity, failing to identify bacteria in four cases that were positive by both BR-PCR and culture.
- While BR-PCR matched culture in detecting bacterial presence, its accuracy in identifying specific bacterial species was inferior to culture methods.
Conclusions:
- Broad-range PCR offers comparable diagnostic accuracy to culture for detecting bacteria in prosthetic joint infections.
- Current molecular identification techniques, including direct sequencing and TaqMan assays, require further refinement to match the species-level identification capabilities of culture.
- Advancements in molecular methods are crucial for improving the identification of causative microorganisms in PJI diagnostics.
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