Abstract
BackgroundOsteomyelitis is a devastating condition whose treatment relies on the detection of bacteria. The current standard of microbiology culture may not be adequate. Molecular biology based diagnostic procedures for detecting bacteria in orthopaedic infections was previously established, but has not been applied to the setting of chronic osteomyelitis. We aim to determine the applicability of molecular diagnostic procedures for monitoring chronic osteomyelitis, and to evaluate if these procedures are superior to standard culture methods of osteomyelitis detection.MethodsA rabbit experimental model of chronic osteomyelitis was used; infection was induced in the proximal, medial aspect of the tibia with Staphylococcus aureus at titers ranging from 1 × 102 to 1 × 106 colony forming units. At 28 days post-infection, animals were sacrificed, and the tibias were examined radiographically, harvested, and assayed for the presence of bacteria. Two bacterial detection methods were used: (1) standard microbiological culturing, and (2) polymerase chain reaction (PCR) based diagnostic method to detect bacterial genomic DNA.ResultsThe molecular diagnostic method was highly sensitive and accurate, and detected low titer infections that were undetected by radiographic and microbiological methods. By using two sets of PCR primers, one for a universal bacterial gene (16S rRNA) and one for a species-specific gene (nuc), the molecular protocol allowed both the detection and speciation of the bacterial infection.ConclusionsThe use of the PCR-based method was effective for high-sensitivity detection and identification of bacteria associated with chronic osteomyelitis in a rabbit model. Our findings illustrate the applicability of PCR for monitoring chronic osteomyelitis, which may be useful for improved detection of osteomyelitis organisms in humans.
Highlights
Osteomyelitis is a devastating condition whose treatment relies on the detection of bacteria
We addressed the following questions: (1) Is the molecular diagnostic procedure applicable for monitoring chronic osteomyelitis?; and (2) Does molecular diagnostic perform better than culture methods for osteomyelitis detection?
For the remaining experimental animals (Group 2) that were tested for infection using the four biopsy method, 4 of the 8 rabbits produced at least 1 microbiological culture test that was positive for bacterial growth, but none tested positive for all four biopsy methods (Table 2)
Summary
Osteomyelitis is a devastating condition whose treatment relies on the detection of bacteria. Molecular biology based diagnostic procedures for detecting bacteria in orthopaedic infections was previously established, but has not been applied to the setting of chronic osteomyelitis. We aim to determine the applicability of molecular diagnostic procedures for monitoring chronic osteomyelitis, and to evaluate if these procedures are superior to standard culture methods of osteomyelitis detection. Enhanced diagnostics for infectious diseases could play a significant role in treating infections, especially in cases where the diagnosis of infection is not straightforward, such as fracture nonunions [21], painful prosthetic joints (Mariani et al 1996b), loose prosthetic joints [22], certain arthritic conditions [23], and osteomyelitis subsequent to bone trauma [24] or as a result of secondary infection from hematogenous sources
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