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Investigation of Biofilm Formation in Clinical Isolates of Staphylococcus aureus

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1085)

Abstract

Invasive methicillin-resistant Staphylococcus aureus (MRSA) infections are often characterized by recalcitrance to antimicrobial therapy, which is a function not only of widespread antimicrobial resistance among clinical isolates, but also the capacity to form biofilms. Biofilms consist of ordered populations of bacterial colonies encased in a polysaccharide and/or proteinaceous matrix. This unique physiologic adaptation limits penetration of antimicrobial molecules and innate immune effectors to the infectious focus, increasing the likelihood of treatment failure and progression to chronic infection. Investigation of mechanisms of biofilm formation and dispersal, as well as the physiologic adaptations to the biofilm lifestyle, is therefore critical to developing new therapies to combat MRSA infections. In this chapter, we describe two in vitro methods for the investigation of staphylococcal biofilm formation, a microtiter plate-based assay of biofilm formation under static conditions and a flow cell-based assay of biofilm formation under fluid shear. We also detail an in vivo murine model of catheter-associated biofilm formation that is amenable to imaging and microbiologic analyses. Special consideration is given to the conditions necessary to support biofilm formation by clinical isolates of S. aureus.

Key words

Polysaccharide intercellular adhesion Poly-N-acetylglucosamine Microbial surface components recognizing adhesive matrix molecules Flow cell Implant-associated biofilm S. aureus MRSA 
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Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  1. 1.Division of Pediatric Infectious DiseasesVanderbilt University Medical CenterNashvilleUSA
  2. 2.Department of Microbiology and ImmunologyUniversity of Arkansas for Medical SciencesLittle RockUSA

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