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Isolation of Chromosomal and Plasmid DNA from Staphylococcus epidermidis

The following describes noncommercial methods for the purification of genomic and plasmid DNA from S. epidermidis. These include both large-scale, high molecular weight and quick, small-scale chromosomal DNA extractions, and also a standard alkaline lysis method of plasmid prepa ...

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Genetic Manipulation of Staphylococci

The ability to genetically manipulate bacteria is essential to understanding gene/protein function in these organisms. While basic cloning has become routine in molecular biology, many still view the ability to make directed mutations as a daunting or intimidating task. To aid the stap ...

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The Isolation and Analysis of Phenol-Soluble Modulins of Staphylococcus epidermidis

Phenol-soluble modulins (PSMs) are multifunctional peptide toxins produced by many staphylococcal strains. PSMs have received much recent attention, owing to multiple reports underscoring their importance for staphylococcal pathogenesis. Members of the PSM family may be s ...

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Growth and Preparation of Staphylococcus epidermidis for NMR Metabolomic Analysis

The “omics” era began with transcriptomics and this progressed into proteomics. While useful, these approaches provide only circumstantial information about carbon flow, metabolic status, redox poise, etc. To more directly address these metabolic concerns, researchers have tu ...

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Multilocus Sequence Typing of Staphylococcus epidermidis

Multilocus sequence typing (MLST) is a genotyping method that is well suited for studying the population genetics and evolution of Staphylococcus epidermidis. The central MLST database for S. epidermidis continues to grow, and new analysis methods for extracting historical inform ...

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Pulsed Field Gel Electrophoresis of Staphylococcus epidermidis

Pulsed field gel electrophoresis (PFGE) is one of the older methods for the molecular characterization and comparison of microorganisms including bacteria. Nevertheless, PFGE continues to be recognized as the gold standard for molecular typing due to output spanning 90 % of the bacter ...

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Identification of Staphylococcus epidermidis in the Clinical Microbiology Laboratory by Molecular Methods

Biochemical assays for the phenotypic identification of coagulase-negative staphylococci in the clinical microbiology laboratory have been well described in previous publications (Becker and Von Eiff Manual of Clinical Microbiology, ASM Press, Washington, pp. 308–330, 20 ...

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Staphylococcus epidermidis Pathogenesis

Staphylococcus epidermidis is the most frequently encountered member of the coagulase-negative staphylococci on human epithelial surfaces. It has emerged as an important nosocomial pathogen, especially in infections of indwelling medical devices. The mechanisms that S. ep ...

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Clinical Characteristics of Infections in Humans Due to Staphylococcus epidermidis

Staphylococcus epidermidis is the most common cause of primary bacteremia and infections of indwelling medical devices. The ability to cause disease is linked to its natural niche on human skin and ability to attach and form biofilm on foreign bodies. This review focuses on the S. epidermidis c ...

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Rat Jugular Catheter Model of Biofilm-Mediated Infection

Staphylococcus epidermidis is now recognized as the primary cause of nosocomial catheter-mediated infections. Bacteria may be introduced exogenously via contamination of the catheter hub or insertion site and endogenously from sepsis. The in vivo model described in this chapter ...

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Generation of a Central Nervous System Catheter-Associated Infection in Mice with Staphylococcus epidermidis

Animal models are valuable tools for investigating the in vivo pathogenesis of Staphylococcus epidermidis infections. Here, we present the procedure for generating a central nervous system catheter-associated infection in a mouse, to model the central nervous system shunt infec ...

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A Mouse Model of Staphylococcus Catheter-Associated Biofilm Infection

Biofilms are adherent communities of bacteria contained within a complex matrix. Staphylococcal species are frequent etiological agents of device-associated biofilm infections in humans that are highly recalcitrant to antimicrobial therapy and alter host immune respons ...

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Mouse Model of Post-arthroplasty Staphylococcus epidermidis Joint Infection

Animal models are invaluable tools for translational research, allowing investigators to recapitulate observed clinical scenarios within the laboratory that share attributes with human disease. Here, we describe a mouse model of post-arthroplasty Staphylococcus epider ...

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Bacteriophage Transduction in Staphylococcus epidermidis

The genetic manipulation of Staphylococcus epidermidis for molecular experimentation has long been an area of difficulty. Many of the traditional laboratory techniques for strain construction are laborious and hampered by poor efficiency. The ability to move chromosomal gene ...

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Rapid Quantitative and Qualitative Analysis of Biofilm Production by Staphylococcus epidermidis Under Static Growth Conditions

Rapid screening of biofilm forming capacity by Staphylococcus epidermidis is possible using in vitro assays with 96-well plates. This method first developed by Christensen et al. in 1985 is fast and does not require specialized instruments. Thus, laboratories with standard microbio ...

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Examination of Staphylococcus epidermidis Biofilms Using Flow-Cell Technology

A common in vitro method to study Staphylococcus epidermidis biofilm development is to allow the bacteria to attach and grow on a solid surface in the presence of a continuous flow of nutrients. Under these conditions, the bacteria progress through a series of developmental steps, ultimately ...

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Methods to Generate a Sequence-Defined Transposon Mutant Library in Staphylococcus epidermidis Strain 1457

Transposon mutant libraries are valuable resources to investigators studying bacterial species, including Staphylococcus epidermidis, which are difficult to genetically manipulate. Although sequence-defined transposon mutant libraries have been constructed in ...

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Use of Electroporation and Conjugative Mobilization for Genetic Manipulation of Staphylococcus epidermidis

To perform mechanistic studies on the biology of bacteria including metabolism, physiology, and pathogenesis, it is essential to possess the tools required for genetic manipulation. Introduction of plasmid DNA into Staphylococcus epidermidis for subsequent genetic manipul ...

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Isolation of Staphylococcus sp. RNA

Isolation of RNA (ribonucleic acid) is a valuable technique to study gene regulation and functional RNAs. It is important to obtain pure samples of RNA for downstream applications, while avoiding the negative effects of ribonucleases (RNases). Here we describe several methods of extrac ...

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Purification, Characterization, and Biotinylation of Single-Chain Antibodies

The variable region (Fv) portion of an antibody is comprised of the antibody VH and VL domains and is the smallest antibody fragment containing a complete antigen-binding site. To stabilize the association of the recombinant VH and VL domains, they have been linked in single-chain Fv constructs ...

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