Characterization of a mixed MRSA/MRSE biofilm in an explanted total ankle arthroplasty

Stoodley, Paul, Conti, Stephen F., DeMeo, Patrick J., Nistico, Laura, Melton-Kreft, Rachael, Johnson, Sandra, Darabi, Ali, Ehrlich, Garth D., Costerton, J. William and Kathju, Sandeep (2011) Characterization of a mixed MRSA/MRSE biofilm in an explanted total ankle arthroplasty. FEMS Immunology & Medical Microbiology, 61 (1), 66-74. (doi:10.1111/j.1574-695X.2011.00793.x). (PMID:21332826)

Abstract

Bacterial biofilms have been observed in many prosthesis-related infections, and this mode-of-growth renders the infection both difficult to treat, and especially difficult to detect and diagnose by standard culture methods. We (1) tested a novel coupled PCR-mass spectroscopic assay (the Ibis T5000) on an ankle arthroplasty that was culture negative on pre-operative aspiration, and then (2) confirmed that the Ibis assay had in fact detected viable multi-species biofilm by further micrographic and molecular examinations, including confocal microscopy using Live/Dead stain, bacterial fluorescent in situ hybridization (FISH), and reverse-transciptase-PCR (RT-PCR) assay for bacterial messenger RNA. The Ibis technology detected Staphylococcus aureus, Staphylococcus epidermidis, and the methicillin resistance gene mecA in soft tissues associated with the explanted hardware. Viable S. aureus were confirmed using RT-PCR, and viable cocci in biofilm configuration were detected microscopically on both tissue and hardware. Species-specific bacterial FISH confirmed a poly-microbial biofilm containing S. aureus. A novel culture method recovered S. aureus and S. epidermidis (both methicillin-resistant) from the tibial metal component. These observations suggest that molecular methods, particularly the new Ibis methodology, may be a useful adjunct to routine cultures in the detection of biofilm bacteria in prosthetic joint infection

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Contributors

Author: Paul Stoodley

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