Table Of ContentMethods in
Molecular Biology 1106
Paul D. Fey Editor
Staphylococcus
Epidermidis
Methods and Protocols
M M B ™
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life Sciences
University of Hertfordshire
Hatfield, Hertfordshire, AL10 9AB, UK
For further volumes:
http://www.springer.com/series/7651
Staphylococcus Epidermidis
Methods and Protocols
Edited by
Paul D. Fey
Department of Pathology and Microbiology, Center for Staphylococcal
Research, University of Nebraska Medical Center, Omaha, NE, USA
Editor
Paul D. Fey
Department of Pathology and Microbiology
Center for Staphylococcal Research
University of Nebraska Medical Center
Omaha , N E, USA
ISSN 1064-3745 ISSN 1940-6029 (electronic)
ISBN 978-1-62703-735-8 ISBN 978-1-62703-736-5 (eBook)
DOI 10.1007/978-1-62703-736-5
Springer New York Heidelberg Dordrecht London
Library of Congress Control Number: 2013953315
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Pref ace
The chapters in this book are designed to give the new investigator a series of tools to ask
novel and exciting questions related to the biology of Staphylococcus epidermidis and other
staphylococci. We are all indebted to the original pioneers (G. Pulverer, G. Peters, F. Götz,
W. Kloos, G. Archer, G. Christensen, K. Schleifer, J. Parisi, and W. Noble among others)
who chose to study this commensal and somewhat recalcitrant opportunistic pathogen.
These original studies, which primarily focused on the establishment of S . epidermidis as an
opportunistic pathogen, laid the groundwork for our current understanding of the patho-
genesis of S . epidermidis including biofi lm formation. However, many exciting and unex-
plored questions remain, such as defi ning the interaction of S . epidermidis and other normal
fl ora. Does S . e pidermidis have a function in immune system development? Why do certain
coagulase negative staphylococci have specifi c microniches on the human host? What fac-
tors function to mediate the binding of S . epidermidis to epithelial cells? Due to the highly
collaborative nature of investigators working in the fi eld, we have rapidly advanced our
understanding of this opportunistic pathogen in the last 20 years. However, due to the
commensal relationship between S . epidermidis and the human host, I predict that the next
20 years will bring highly novel insights regarding the function of S . epidermidis coloniza-
tion on human development and overall health.
Omaha, NE, USA Paul D. Fey
May 2013
v
Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
1 Clinical Characteristics of Infections in Humans
Due to Staphylococcus epidermidis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Mark E. Rupp
2 Staphylococcus epidermidis Pathogenesis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Michael Otto
3 Identification of Staphylococcus epidermidis in the Clinical Microbiology
Laboratory by Molecular Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
Amity L. Roberts
4 Pulsed Field Gel Electrophoresis of Staphylococcus epidermidis. . . . . . . . . . . . . . . . . . 55
Richard V. Goering and Paul D. Fey
5 Multilocus Sequence Typing of Staphylococcus epidermidis . . . . . . . . . . . . . . . . . . . . . 61
Jonathan C. Thomas and D. Ashley Robinson
6 Growth and Preparation of Staphylococcus epidermidis
for NMR Metabolomic Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
Greg A. Somerville and Robert Powers
7 The Isolation and Analysis of Phenol-Soluble Modulins
of Staphylococcus epidermidis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
Hwang-Soo Joo and Michael Otto
8 Genetic Manipulation of Staphylococci. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101
Jeffrey L. Bose
9 Isolation of Chromosomal and Plasmid DNA from Staphylococcus epidermidis . . . . . 113
Jill K. Lindgren
10 Isolation of Staphylococcus sp. RNA. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
Tess Eidem
11 Use of Electroporation and Conjugative Mobilization
for Genetic Manipulation of Staphylococcus epidermidis. . . . . . . . . . . . . . . . . . . . . . . . 125
Katherine L. Maliszewski and Austin S. Nuxoll
12 Methods to Generate a Sequence-Defined Transposon Mutant
Library in Staphylococcus epidermidis Strain 1457. . . . . . . . . . . . . . . . . . . . . . . . . . . 135
Todd J. Widhelm, Vijay Kumar Yajjala, Jennifer L. Endres,
Paul D. Fey, and Kenneth W. Bayles
13 Examination of Staphylococcus epidermidis Biofilms
Using Flow-C ell Technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143
Derek E. Moormeier and Kenneth W. Bayles
vii
viii Contents
14 Rapid Quantitative and Qualitative Analysis of Biofilm Production
by Staphylococcus epidermidis Under Static Growth Conditions . . . . . . . . . . . . . . . . 157
Elaine M. Waters, Hannah McCarthy, Siobhan Hogan,
Marta Zapotoczna, Eoghan O’Neill, and James P. O’Gara
15 Bacteriophage Transduction in Staphylococcus epidermidis. . . . . . . . . . . . . . . . . . . . . . 167
Michael E. Olson and Alexander R. Horswill
16 Mouse Model of Post-arthroplasty Staphylococcus epidermidis Joint Infection. . . . . . 173
Tyler D. Scherr, Kevin E. Lindgren, Carolyn R. Schaeffer,
Mark L. Hanke, Curtis W. Hartman, and Tammy Kielian
17 A Mouse Model of Staphylococcus Catheter-Associated Biofilm Infection. . . . . . . . . 183
Cortney E. Heim, Mark L. Hanke, and Tammy Kielian
18 Generation of a Central Nervous System Catheter-A ssociated Infection in Mice
with Staphylococcus epidermidis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 193
Jessica N. Snowden
19 Rat Jugular Catheter Model of Biofilm-Mediated Infection. . . . . . . . . . . . . . . . . . . 199
Carolyn R. Schaeffer, Keith M. Woods, and G. Matthew Longo
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207
Contributors
KENNETH W. BAYLES ∙ Department of Pathology and Microbiology, Center for
Staphylococcal Research, University of Nebraska Medical Center , Omaha , NE , USA
JEFFREY L. BOSE ∙ Department of Pathology and Microbiology, Center for Staphylococcal
Research, University of Nebraska Medical Center , Omaha , NE , USA
TESS EIDEM ∙ Department of Pathology and Microbiology, University of Nebraska
Medical Center , Omaha , NE , USA
JENNIFER L. ENDRES ∙ Department of Pathology and Microbiology, Center for
Staphylococcal Research, University of Nebraska Medical Center , Omaha , NE , USA
PAUL D. FEY ∙ Department of Pathology and Microbiology, Center for Staphylococcal
Research, University of Nebraska Medical Center , Omaha , NE , USA
RICHARD V. GOERING ∙ Department of Medical Microbiology and Immunology,
Creighton University School of Medicine , Omaha , NE , USA
MARK L. HANKE ∙ Department of Pathology and Microbiology, Center for Staphylococcal
Research, University of Nebraska Medical Center , Omaha , NE , USA
CURTIS W. HARTMAN ∙ Department of Orthopedic Surgery , University of Nebraska
Medical Center , Omaha , NE , USA
CORTNEY E. HEIM ∙ Department of Pathology and Microbiology, Center for
Staphylococcal Research, University of Nebraska Medical Center , Omaha , NE , USA
SIOBHAN HOGAN ∙ Department of Clinical Microbiology, Royal College of Surgeons in
Ireland , Dublin, Ireland
ALEXANDER R. HORSWILL ∙ Department of Microbiology, University of Iowa , Iowa City,
IA , USA
HWANG-SOO JOO ∙ Pathogen Molecular Genetics Section, Laboratory of Human
Bacterial Pathogenesis, National Institute of Allergy and Infectious Diseases, U.S.
National Institutes of Health , Bethesda , MD , USA
TAMMY KIELIAN ∙ Department of Pathology and Microbiology, Center for Staphylococcal
Research, University of Nebraska Medical Center , Omaha , NE , USA
KEVIN E. LINDGREN ∙ Department of Orthopedic Surgery , University of Nebraska
Medical Center , Omaha , NE , USA
JILL K. LINDGREN ∙ Department of Pathology and Microbiology, Center for
Staphylococcal Research, University of Nebraska Medical Center , Omaha , NE , USA
G. MATTHEW LONGO ∙ Division of Vascular Surgery, Department of General Surgery ,
University of Nebraska Medical Center , Omaha , NE , USA
KATHERINE L. MALISZEWSKI ∙ Department of Pathology and Microbiology, Center for
Staphylococcal Research, University of Nebraska Medical Center , Omaha , NE , USA
HANNAH MCCARTHY ∙ Microbiology, School of Natural Sciences, National University
of Ireland , Galway, Ireland
DEREK E. MOORMEIER ∙ Department of Pathology and Microbiology, Center for
Staphylococcal Research, University of Nebraska Medical Center , Omaha , NE , USA
ix
