Authors of previous studies have found that culture-based detection of prosthetic hip infections can be improved by adherence to strict anaerobic protocols and mild ultrasonication of retrieved prostheses to remove bacteria growing as an adherent biofilm. Furthermore, direct analyses of sonicate samples by immunofluorescence microscopy (with genus-specific and species-specific monoclonal antibodies or polyclonal antisera) produces significantly greater and more rapid detection rates for infection compared with culture alone. Despite its diagnostic value within a research setting, the practical advantages and limitations of immunofluorescence microscopy for a busy diagnostic laboratory needs consideration. In addition, the method must be evaluated against other nonculture-based techniques that may prove more appropriate. In this review, we describe how immunofluorescence microscopy presents an attractive and reliable method for routine detection of prosthetic hip biofilms compared, primarily, to polymerase chain reaction, although other nonculture methods of diagnosis based on fluorescent in situ hybridization and serologic analysis are considered. We describe how the immunofluorescence microscopy technique is robust, relatively simple and, in contrast to broad-range 16S recombinant-deoxyribonucleic-acid-based polymerase chain reaction, does not require a separate clean-room facility or strict adherence to aseptic techniques. More importantly we highlight how dislodged biofilm, which appears as large aggregates of bacteria, easily can be identified from skin contaminants that occur as single cells or small aggregates of only a few cells.

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