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Fast and Sensitive Multiplex Real-Time Quantitative PCR to Detect Cutibacterium Periprosthetic Joint Infections
Julia Prinz1, Bettina Schmid2, Reinhard Zbinden3
1Department of Dermatology, University Hospital Zurich, University of Zurich, Zurich, Switzerland; Department of Infectious Diseases and Hospital Epidemiology, University Hospital Zurich, University of Zurich, Zurich, Switzerland.
Abstract:
Diagnosis of Cutibacterium periprosthetic joint infections (PJIs) is challenging due to a long cultivation time of up to 14 days. Faster culture-independent diagnosis would improve patient care with early and accurate treatment. Specific primers and probes were designed for Cutibacterium acnes, Cutibacterium avidum, and Cutibacterium granulosum and evaluated in a multiplex TaqMan real-time quantitative PCR (qPCR) format on 57 skin swabs and 20 culture-negative cerebrospinal fluid samples. The multiplex qPCR was tested in a PJI cohort of 41 sonication fluid samples from removed implants infected with different pathogens. All five culture-positive Cutibacterium PJIs were detected with the corresponding Cutibacterium-specific probe (100% positive percent agreement). The multiplex qPCR additionally detected C. avidum in two PJI sonication fluid samples that were diagnosed as Staphylococcus species infections according to culture (95% negative percent agreement). The new multiplex qPCR can provide a Cutibacterium PJI diagnosis within 1 day, allowing early and accurate antibiotic treatment. A prospective diagnostic trial in PJI with a high number of Cutibacterium species infections (shoulder PJI) is needed for further evaluation.
Insights
A new multiplex qPCR test offers rapid diagnosis of Cutibacterium periprosthetic joint infections (PJIs) within one day. This faster method aids early and accurate antibiotic treatment for patients with these challenging infections.
Area of Science:
- Microbiology
- Infectious Diseases
- Molecular Diagnostics
Background:
- Periprosthetic joint infections (PJIs) caused by Cutibacterium species present diagnostic challenges due to lengthy culture times (up to 14 days).
- Delayed diagnosis of Cutibacterium PJIs can impede timely and effective patient treatment.
- Culture-independent methods are needed for faster and more accurate PJI diagnostics.
Purpose of the Study:
- To develop and evaluate a multiplex real-time quantitative PCR (qPCR) assay for the rapid detection of Cutibacterium species in PJI samples.
- To assess the diagnostic performance of the multiplex qPCR compared to traditional culture methods.
Main Methods:
- Design of specific primers and probes for Cutibacterium acnes, C. avidum, and C. granulosum.
- Development of a multiplex TaqMan real-time quantitative PCR (qPCR) assay.
- Validation using skin swabs, cerebrospinal fluid samples, and sonication fluid from PJI patients.
Main Results:
- The multiplex qPCR achieved 100% positive percent agreement for culture-positive Cutibacterium PJIs.
- It detected C. avidum in two PJI samples misidentified by culture as Staphylococcus species (95% negative percent agreement).
- The assay provides diagnostic results within one day, significantly faster than conventional cultures.
Conclusions:
- The developed multiplex qPCR assay enables rapid and accurate diagnosis of Cutibacterium PJIs.
- This molecular approach facilitates early initiation of appropriate antibiotic therapy.
- Further prospective trials, particularly in high Cutibacterium prevalence settings like shoulder PJIs, are warranted for full evaluation.
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