Staphylococcus aureus Methods and Protocols

This volume details the phenotypic characterization of Staphylococcus aureus, with a focus on in vitro and ex vivo methodologies. The chapters in this book cover topics such as in vitro assessment of classical S. aureus virulence attributes; quantifying promoter activity using a S. aureus codon-optimized lacZ plasmid; biologically-relevant growth environments; metabolic and stress resistance assays; and in vivo and ex vivo models of host-pathogen interaction. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Preface…

Table of Contents…

Contributing Authors…

1. Measurement of Staphylococcus aureus Pigmentation by Methanol Extraction

Leia E. Sullivan and Kelly C. Rice

2. Analysis of Murein Hydrolases and Proteases through Zymography

Nichole A. Seawell and Jeffrey L. Bose

3. Detection and Quantification of Secreted Nuclease Activity in Staphylococcus aureus Culture Supernatants

Richard E. Wiemels, Rebecca A. Keogh, and Ronan K. Carroll

4. Quantitative Hemolysis Assays

Miranda J. Ridder, Seth M. Daly, Pamela R. Hall, and Jeffrey L. Bose

5. In Vitro Assay for Quantifying Clumping of Staphylococcus aureus 

Heidi A. Crosby, Jakub M. Kwiecinski, and Alexander R. Horswill

6. Measuring Staphylococcal Promoter Activities using a Codon-Optimized ß-galactosidase Reporter

7. Evaluation of Staphylococcus aureus Antibiotic Tolerance using Kill Curve Assays

Jessica N. Brandwein and Kelly C. Rice

8. Fluorescence Polarization (FP) Assay for Measuring Staphylococcus aureus Membrane Fluidity

Kiran B. Tiwari, Suranjana Sen, Craig Gatto, and Brian J. Wilkinson

9. Quantification of Staphylococcus aureus Biofilm Formation by Crystal 

Violet and Confocal Microscopy 

Adam B. Grossman, Dylan J. Burgin, and Kelly C. Rice

10. Growth of Staphylococcus aureus using a Rotary Cell Culture System 

Matthew R. Hauserman and Kelly C. Rice 

11. Time-Resolved Fluorescence Assay for Measuring Oxygen Consumption Rates in Staphylococcus aureus

Logan L. Bulock, Jennifer L. Endres, and Marat R. Sadykov

12. Quantifying Staphylococcus aureus Membrane Potential using Flow Cytometry

Neal D. Hammer

13. Best Practices for Preparation of Staphylococcus aureus Metabolomics Samples 

Kimberly L. James and Kelly C. Rice

14. A Rat Model of Orthopedic Implant-Associated Infection for Identification of Staphylococcal Biofilm Proteins

Mei G. Lei, Ravi Kr. Gupta, and Chia Y. Lee

15. An Ex Vivo Model for Assessing Growth and Survivability of Staphylococcus aureus in Whole Human Blood 

Brittney D. Gimza, Stephanie M. Marroquin, and Lindsey N. Shaw

16. Bone Marrow-Derived Macrophage Infection Assay

Miranda J. Ridder, Mary A. Markiewicz, and Jeffrey L. Bose

17. Measurement of Osteoblast Cytotoxicity Induced by S. aureus Secreted Toxins

Caleb Ford and James E. Cassat

18. Human Skin In Vitro Colonization Model for a Skin Wound Infected by Staphylococcus aureus Biofilm

Jenelle Chapman and Michael E. Olson

Subject Index List…