Colonization of surfaces in the human body by microorganisms is an early, essential, step in the initiation of infectious disease. We have developed in vitro assays to investigate interactions between yeast or bacterial cells and human tissues, fluids, or prostheses. Such assays can be used to identify the adhesins, ligands, and receptors involved in these interactions, for example, by determining which components of the microbe or human tissue/fluid interfere with adherence in the assay. The assays can also be applied to finding ways of preventing adhesion, and subsequent disease, by investigating the effects of different conditions and added compounds on adherence in the in vitro assays.
Here we describe assays for measuring adhesion of the oral yeast Candida albicans, a common commensal and opportunistic pathogen, or the bacterium Staphylococcus epidermidis, which is not normally pathogenic but is known to form biofilms on medical prostheses. The assays described belong to two approaches to investigating adhesion and biofilm formation: (1) retention at a fixed time point following liquid washes and (2) retention against a continuous flow of medium.
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We are grateful to Andrew McNaughton (Otago Centre for Confocal Microscopy) for undertaking the confocal microscopy analysis and to Steve Swindells (University of Otago School of Dentistry) for the assistance with the fabrication of the acrylic templates used in the biofilm assay. We thank Professor Henk Busscher (Department of Biomedical Engineering, University of Groningen, the Netherlands) for the advice on the parallel plate flow apparatus. We gratefully acknowledge funding from the New Zealand Lottery Grants Board, the New Zealand Dental Association Research Foundation, and the University of Otago. K. N.-W. is grateful to the Sir John Walsh Research Institute for the award of a Fuller Scholarship.
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