Center for Medical Mycology and Department of Dermatology, University Hospitals Case Medical Center and Case Western Reserve University, Cleveland, OH, USA.
Virulence. 2011 Sep-Oct;2(5):466-74. doi: 10.4161/viru.2.5.16341. Epub 2011 Sep 1.
Most cases of catheter-related bloodstream infections (CRBSIs) involve colonization of micro-organisms on catheter surfaces where they eventually become embedded in a biofilm. Fungal biofilm formation is studied using a number of techniques, involving the use of a wide variety of substrates and growth conditions. In vitro techniques involving use of confocal scanning laser/scanning electron microscopy, metabolic activity assay, dry weight measurements and antifungal susceptibility assays are increasingly used by investigators to quantify and evaluate biofilm morphology. However, there are not many in vivo models used to validate biofilm-associated infections. In this protocol, we describe clinically relevant rabbit model of C. albicans biofilm-associated catheter infection to evaluate the morphology, topography, and architecture of fungal biofilms. The methods described here can be completed in a typical laboratory setting. Evaluation of the formation of fungal biofilms on catheters in vivo, their analysis using scanning electron microscopy (SEM) and quantitative catheter culture (QCC) and treatment of biofilms using antimicrobial lock therapy can be completed using the described methods in ~20-25 days. This model has utility in evaluating the efficacy of lock solutions. In addition, it is a useful approach for characterizing/comparing the formation of biofilms by wild type and isogenic mutants including clinical isolates in vivo. This model can also be used for testing different biomaterials.
大多数导管相关血流感染(CRBSI)病例涉及微生物在导管表面的定植,最终它们会嵌入生物膜中。真菌生物膜形成的研究使用了多种技术,涉及到使用各种不同的基质和生长条件。研究人员越来越多地使用涉及共聚焦扫描激光/扫描电子显微镜、代谢活性测定、干重测量和抗真菌药敏试验的体外技术来定量和评估生物膜形态。然而,用于验证生物膜相关感染的体内模型并不多。在本方案中,我们描述了一种临床相关的白色念珠菌生物膜相关导管感染兔模型,用于评估真菌生物膜的形态、形貌和结构。这里描述的方法可以在典型的实验室环境中完成。在体内评估导管上真菌生物膜的形成,使用扫描电子显微镜(SEM)和定量导管培养(QCC)进行分析,以及使用抗菌封管治疗生物膜,可以使用所述方法在 20-25 天内完成。该模型可用于评估封管溶液的疗效。此外,它是一种在体内评估野生型和同源突变体(包括临床分离株)生物膜形成的有用方法。该模型还可用于测试不同的生物材料。