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Updated: May 12, 2025

Novel Diagnostics in Revision Arthroplasty: Implant Sonication and Multiplex Polymerase Chain Reaction
Published on: December 3, 2017
Multi-centre evaluation of Gram stain in the diagnosis of septic arthritis
Charlotte Smith1,1, Robert J Maloney2,2, Deborah Wearmouth2,2
1Sheffield Health & Social Care NHS Foundation Trust, Centre Court, Atlas Way, Sheffield, S4 7QQ, UK.
Abstract:
Introduction: Gram stain of synovial fluid is a rapid test for the diagnosis of native joint septic arthritis. Single-centre studies have suggested Gram stain will miss a considerable proportion of patients who are subsequently synovial-fluid-culture-positive or polymerase chain reaction (PCR)-positive. The object of this study was to reassess Gram stain in a large, multi-centre cohort of patients from the United Kingdom (UK) and Ireland. Methods: The study was a retrospective analysis combining two large datasets. We defined septic arthritis microbiologically as at least one positive joint aspirate culture and/or PCR test. "Best case" and "worst case" definitions were applied depending on the likelihood organisms were true infecting pathogens. Results: Gram stain missed a high proportion of culture-/PCR-positive patients using both the best (74 % missed) and worst (81 % missed) case definitions. Using the best case definition, the sensitivity of Gram stain was 0.26, specificity 0.99, positive predictive value 0.84, negative predictive value 0.87, accuracy 0.87, and area under the receiver operator curve 0.62 (95 % CI 0.57 to 0.68, ). False positive Gram stains were infrequent (1 %). Age, joint involved, and other synovial fluid characteristics were less predictive of a positive culture/PCR than Gram stain. Conclusions: While a positive synovial fluid Gram stain should always be considered to indicate potential septic arthritis, a negative Gram stain, regardless of synovial fluid crystals or white cell count, should not be used to rule out septic arthritis. The value of Gram stain as an urgent out-of-hours test for septic arthritis is open to considerable debate.
Insights
Gram stain of synovial fluid is a poor test for diagnosing septic arthritis, frequently missing infections confirmed by culture or PCR. A negative Gram stain should not rule out septic arthritis, even with other abnormal synovial fluid findings.
Area of Science:
- Orthopedics
- Infectious Diseases
- Diagnostic Microbiology
Background:
- Synovial fluid Gram stain is a rapid diagnostic test for native joint septic arthritis.
- Previous single-centre studies indicated Gram stain misses a significant number of culture- or PCR-positive septic arthritis cases.
- The diagnostic utility of Gram stain for septic arthritis requires re-evaluation in a larger, multi-centre setting.
Purpose of the Study:
- To reassess the accuracy of synovial fluid Gram stain in diagnosing septic arthritis.
- To evaluate the sensitivity and specificity of Gram stain in a large, multi-centre UK and Ireland cohort.
- To determine the role of Gram stain in the urgent diagnosis of septic arthritis.
Main Methods:
- Retrospective analysis of two large datasets from the UK and Ireland.
- Defined septic arthritis microbiologically by positive joint aspirate culture and/or PCR.
- Applied "best case" and "worst case" definitions to assess Gram stain performance.
Main Results:
- Gram stain missed a high proportion of culture-/PCR-positive septic arthritis cases (74-81%).
- Sensitivity was low (0.26) with high specificity (0.99) using the "best case" definition.
- False positive Gram stains were rare (1%), and other synovial fluid parameters were less predictive than Gram stain.
Conclusions:
- A positive synovial fluid Gram stain is indicative of potential septic arthritis.
- A negative Gram stain should not exclude septic arthritis, irrespective of synovial fluid crystals or white cell count.
- The value of Gram stain as an urgent diagnostic test for septic arthritis is questionable.

