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Fluorescence “in situ” hybridization for the detection of biofilm in the middle ear and upper respiratory tract mucosa

Fluorescence “in situ” hybridization for the detection of biofilm in the middle ear and upper respiratory tract mucosa
Fluorescence “in situ” hybridization for the detection of biofilm in the middle ear and upper respiratory tract mucosa
Most chronic bacterial infections are associated with biofilm formation wherein the bacteria attach to mucosal surfaces, wound tissue, or medical device surfaces in the human body via the formation of an extracellular matrix. Biofilms assume complex three-dimensional structures dependent on the species, the strain, and the prevailing environmental conditions and are composed of both the bacteria and the extracellular slime-like matrices, which surround the bacteria. Bacteria deep in the biofilm live under anaerobic conditions and must use alternatives to O(2) as a terminal electron acceptor. Thus, the metabolic rates of these deep bacteria are greatly reduced, which renders them extremely resistant to antibiotic treatment, and for reasons not clearly understood, it is often very difficult to culture biofilm bacteria using traditional microbiologic techniques. To directly identify and visualize biofilm bacteria in a species-specific manner, we developed a confocal laser scanning microscopy (CLSM)-based 16S rRNA fluorescence in situ hybridization (FISH) protocol, to find biofilm bacteria in middle ear and upper respiratory tract mucosa, which preserves the three-dimensional structure of the biofilm and avoids the use of traditional culture techniques.
middle ear mucosa, upper respiratory tract mucosa, biofilm, FISH, 16S rRNA fluorescent probes, confocal laser scanning microscopy (CLSM)
9781934115626
1064-3745
493
191-213
Springer
Nistico, Laura
7a83886a-6bf1-46a1-87dd-75a120d41603
Gieseke, Armin
1b9305ea-9172-4681-8667-9f638aac41f2
Stoodley, Paul
08614665-92a9-4466-806e-20c6daeb483f
Hall-Stoodley, Luanne
94ebdc00-b549-4488-b15f-5310fb965f5b
Kerschner, Joseph E.
802c8916-fcb7-4f89-8555-6643935735ec
Ehrlich, Garth D.
aa8e5162-77a6-4627-a793-acd724ed0782
Sokolowski, Bernd
Nistico, Laura
7a83886a-6bf1-46a1-87dd-75a120d41603
Gieseke, Armin
1b9305ea-9172-4681-8667-9f638aac41f2
Stoodley, Paul
08614665-92a9-4466-806e-20c6daeb483f
Hall-Stoodley, Luanne
94ebdc00-b549-4488-b15f-5310fb965f5b
Kerschner, Joseph E.
802c8916-fcb7-4f89-8555-6643935735ec
Ehrlich, Garth D.
aa8e5162-77a6-4627-a793-acd724ed0782
Sokolowski, Bernd

Nistico, Laura, Gieseke, Armin, Stoodley, Paul, Hall-Stoodley, Luanne, Kerschner, Joseph E. and Ehrlich, Garth D. (2009) Fluorescence “in situ” hybridization for the detection of biofilm in the middle ear and upper respiratory tract mucosa. In, Sokolowski, Bernd (ed.) Auditory and Vestibular Research: Methods and Protocols. (Methods in Molecular Biology, 493, 493) UK. Springer, pp. 191-213. (doi:10.1007/978-1-59745-523-7_12).

Record type: Book Section

Abstract

Most chronic bacterial infections are associated with biofilm formation wherein the bacteria attach to mucosal surfaces, wound tissue, or medical device surfaces in the human body via the formation of an extracellular matrix. Biofilms assume complex three-dimensional structures dependent on the species, the strain, and the prevailing environmental conditions and are composed of both the bacteria and the extracellular slime-like matrices, which surround the bacteria. Bacteria deep in the biofilm live under anaerobic conditions and must use alternatives to O(2) as a terminal electron acceptor. Thus, the metabolic rates of these deep bacteria are greatly reduced, which renders them extremely resistant to antibiotic treatment, and for reasons not clearly understood, it is often very difficult to culture biofilm bacteria using traditional microbiologic techniques. To directly identify and visualize biofilm bacteria in a species-specific manner, we developed a confocal laser scanning microscopy (CLSM)-based 16S rRNA fluorescence in situ hybridization (FISH) protocol, to find biofilm bacteria in middle ear and upper respiratory tract mucosa, which preserves the three-dimensional structure of the biofilm and avoids the use of traditional culture techniques.

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More information

Published date: 8 October 2009
Additional Information: The National Centre for Advanced Tribology at Southampton (nCATS)
Keywords: middle ear mucosa, upper respiratory tract mucosa, biofilm, FISH, 16S rRNA fluorescent probes, confocal laser scanning microscopy (CLSM)
Organisations: Biological Sciences, Engineering Sciences

Identifiers

Local EPrints ID: 71662
URI: http://eprints.soton.ac.uk/id/eprint/71662
ISBN: 9781934115626
ISSN: 1064-3745
PURE UUID: 3c79e471-bf29-40da-a2b4-7cc44e73de4c
ORCID for Paul Stoodley: ORCID iD orcid.org/0000-0001-6069-273X

Catalogue record

Date deposited: 17 Dec 2009
Last modified: 14 Mar 2024 02:55

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Contributors

Author: Laura Nistico
Author: Armin Gieseke
Author: Paul Stoodley ORCID iD
Author: Luanne Hall-Stoodley
Author: Joseph E. Kerschner
Author: Garth D. Ehrlich
Editor: Bernd Sokolowski

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