Cassat James E, Smeltzer Mark S, Lee Chia Y
Division of Pediatric Infectious Diseases, Vanderbilt University Medical Center, Nashville, TN, USA.
Methods Mol Biol. 2014;1085:195-211. doi: 10.1007/978-1-62703-664-1_12.
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.
侵袭性耐甲氧西林金黄色葡萄球菌（MRSA）感染的特点通常是对抗菌治疗具有顽固性，这不仅是因为临床分离株中广泛存在抗菌耐药性，还因为其形成生物膜的能力。生物膜由包裹在多糖和/或蛋白质基质中的有序细菌菌落群体组成。这种独特的生理适应性限制了抗菌分子和固有免疫效应物渗透到感染灶，增加了治疗失败和进展为慢性感染的可能性。因此，研究生物膜形成和分散的机制以及对生物膜生活方式的生理适应性，对于开发对抗MRSA感染的新疗法至关重要。在本章中，我们描述了两种用于研究葡萄球菌生物膜形成的体外方法，一种是基于微孔板的静态条件下生物膜形成测定法，另一种是基于流动小室的流体剪切条件下生物膜形成测定法。我们还详细介绍了一种适用于成像和微生物学分析的导管相关生物膜形成的体内小鼠模型。特别考虑了支持金黄色葡萄球菌临床分离株形成生物膜所需的条件。
