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Molecular and imaging techniques for bacterial biofilms in joint arthroplasty infections

Molecular and imaging techniques for bacterial biofilms in joint arthroplasty infections
Molecular and imaging techniques for bacterial biofilms in joint arthroplasty infections
Biofilm formation on surfaces is an ancient and integral strategy for bacterial survival. Billions of years of adaptation provide microbes with the ability to colonize any surface, including those used in orthopaedic surgery. Although remarkable progress has been made in the treatment of orthopaedic diseases with implanted prostheses, infection rates remain between 1% and 2%, and are higher for revision surgeries. The chronic nature of implant infections, their nonresponsiveness to antibiotics, and their frequent culture negativity can be explained by the biofilm paradigm of infectious disease. However, the role of biofilms in orthopaedic implant infections and aseptic loosening is controversial. To address these issues, we developed molecular diagnostic and confocal imaging techniques to identify and characterize biofilms associated with infected implants. We designed PCR and reverse transcription (RT)-PCR-based assays that can be used to detect bacterial infections associated with culture-negative joint effusions that distinguish between physiologically active Staphylococcus aureus and Staphylococcus epidermidis. Using clinical isolates of Pseudomonas aeruginosa, we constructed a series of reporter strains expressing colored fluorescent proteins to observe biofilms growing on 316L stainless steel and titanium orthopaedic screws. Three-dimensional structures of Pseudomonas aeruginosa and staphylococci biofilms growing on the screws were documented using confocal microscopy. The application of these tools for clinical diagnosis and biofilm research in animal and in vitro models is discussed.
0009-921X
31-40
Stoodley, Paul
08614665-92a9-4466-806e-20c6daeb483f
Kathju, Sandeep
80cdb7ee-2e0d-4e70-98c9-93682ce05a09
Hu, Fen Ze
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Erdos, Geza
2ce1be9d-2860-4475-a664-012af58a4137
Levenson, Joshua E
16639544-bd4c-4e76-ac58-caa71cfbfab6
Mehta, Nalini
1680b9d7-1639-4a51-9ee1-359fb31bdeff
Dice, Bethany
72120cd5-bde7-4799-bba8-38cc906d14b1
Johnson, Sandy
1b12d9c7-844b-4d38-aefe-d966a2c8aca8
Hall-Stoodley, Luanne
94ebdc00-b549-4488-b15f-5310fb965f5b
Nistico, Laura
7a83886a-6bf1-46a1-87dd-75a120d41603
Sotereanos, Nicholas
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Sewecke, Jeff
4ffbde66-ee85-46da-aa43-32426f840c13
Post, J Christopher
d6d584b1-78bd-44cf-a8f4-255b88ffa904
Ehrlich, Garth D
310053ba-82ce-4a24-94ef-5f3d4ca6f5af
Stoodley, Paul
08614665-92a9-4466-806e-20c6daeb483f
Kathju, Sandeep
80cdb7ee-2e0d-4e70-98c9-93682ce05a09
Hu, Fen Ze
a6e5ffb6-a70d-4a7a-97bc-34b34d5bb0cc
Erdos, Geza
2ce1be9d-2860-4475-a664-012af58a4137
Levenson, Joshua E
16639544-bd4c-4e76-ac58-caa71cfbfab6
Mehta, Nalini
1680b9d7-1639-4a51-9ee1-359fb31bdeff
Dice, Bethany
72120cd5-bde7-4799-bba8-38cc906d14b1
Johnson, Sandy
1b12d9c7-844b-4d38-aefe-d966a2c8aca8
Hall-Stoodley, Luanne
94ebdc00-b549-4488-b15f-5310fb965f5b
Nistico, Laura
7a83886a-6bf1-46a1-87dd-75a120d41603
Sotereanos, Nicholas
dd4dadad-f07a-4bb9-9783-9c7c63bb249f
Sewecke, Jeff
4ffbde66-ee85-46da-aa43-32426f840c13
Post, J Christopher
d6d584b1-78bd-44cf-a8f4-255b88ffa904
Ehrlich, Garth D
310053ba-82ce-4a24-94ef-5f3d4ca6f5af

Stoodley, Paul, Kathju, Sandeep, Hu, Fen Ze, Erdos, Geza, Levenson, Joshua E, Mehta, Nalini, Dice, Bethany, Johnson, Sandy, Hall-Stoodley, Luanne, Nistico, Laura, Sotereanos, Nicholas, Sewecke, Jeff, Post, J Christopher and Ehrlich, Garth D (2005) Molecular and imaging techniques for bacterial biofilms in joint arthroplasty infections. Clinical Orthopaedics and Related Research, (437), 31-40. (doi:10.1097/01.blo.0000175129.83084.d5).

Record type: Article

Abstract

Biofilm formation on surfaces is an ancient and integral strategy for bacterial survival. Billions of years of adaptation provide microbes with the ability to colonize any surface, including those used in orthopaedic surgery. Although remarkable progress has been made in the treatment of orthopaedic diseases with implanted prostheses, infection rates remain between 1% and 2%, and are higher for revision surgeries. The chronic nature of implant infections, their nonresponsiveness to antibiotics, and their frequent culture negativity can be explained by the biofilm paradigm of infectious disease. However, the role of biofilms in orthopaedic implant infections and aseptic loosening is controversial. To address these issues, we developed molecular diagnostic and confocal imaging techniques to identify and characterize biofilms associated with infected implants. We designed PCR and reverse transcription (RT)-PCR-based assays that can be used to detect bacterial infections associated with culture-negative joint effusions that distinguish between physiologically active Staphylococcus aureus and Staphylococcus epidermidis. Using clinical isolates of Pseudomonas aeruginosa, we constructed a series of reporter strains expressing colored fluorescent proteins to observe biofilms growing on 316L stainless steel and titanium orthopaedic screws. Three-dimensional structures of Pseudomonas aeruginosa and staphylococci biofilms growing on the screws were documented using confocal microscopy. The application of these tools for clinical diagnosis and biofilm research in animal and in vitro models is discussed.

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More information

Published date: August 2005
Organisations: Engineering Mats & Surface Engineerg Gp

Identifiers

Local EPrints ID: 155961
URI: http://eprints.soton.ac.uk/id/eprint/155961
ISSN: 0009-921X
PURE UUID: b7b4e894-dcc6-42bd-8121-ca3c191edb8a
ORCID for Paul Stoodley: ORCID iD orcid.org/0000-0001-6069-273X

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Date deposited: 08 Jun 2010 14:17
Last modified: 14 Mar 2024 02:55

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Contributors

Author: Paul Stoodley ORCID iD
Author: Sandeep Kathju
Author: Fen Ze Hu
Author: Geza Erdos
Author: Joshua E Levenson
Author: Nalini Mehta
Author: Bethany Dice
Author: Sandy Johnson
Author: Luanne Hall-Stoodley
Author: Laura Nistico
Author: Nicholas Sotereanos
Author: Jeff Sewecke
Author: J Christopher Post
Author: Garth D Ehrlich

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