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Use of fluorescent in situ hybridisation for the visualisation of Helicobacter pylori in real drinking water biofilms

Use of fluorescent in situ hybridisation for the visualisation of Helicobacter pylori in real drinking water biofilms
Use of fluorescent in situ hybridisation for the visualisation of Helicobacter pylori in real drinking water biofilms
A fluorescently labelled peptide nucleic acid (PNA) probe has been applied for the in situ detection of Helicobacter pylori in drinking water biofilms. The method was originally applied to real pipe samples removed from a drinking water distribution system (DWDS) but the curvature and the heavy fouling of the pipes prevented an accurate detection of the bacterium by epifluorescence microscopy. Therefore, two semi-circular flow cells were placed in a bypass of the DWDS, and coupons with up to 72 days of exposure were regularly sampled and analysed for the presence of H. pylori. In the flat surfaces of the coupons, it was possible to sparsely detect cells exhibiting similar morphology to H. pylori that were emitting the PNA probe fluorescent signal. Coupons were also visualised under the microscope before the hybridisation procedure to serve as negative controls and ensure the validity of the method. This work corroborates the findings already published elsewhere that this bacterium might be present in DWDS biofilms. The method requires, however, highly trained personnel for an accurate detection of the pathogen and will need simplification before being routinely used in standard water analysis laboratories. © IWA Publishing 2007.
Biofilms, Drinking water, Helicobacter pylori, Peptide nucleic acids, Rapid detection methods
0273-1223
387-393
Bragança, Sofia M.
7df84b21-9316-4ee2-9445-4746df073fd8
Azevedo, Nuno F.
79da2c36-313d-41a4-84a1-caaa3f6b0264
Simöes, Lucia Chaves
9dd958fa-f504-4849-9975-fedfc3e9a013
Keevil, C. W.
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Vieira, M. J.
45deadf9-bf20-4e60-95e2-506226f0fed2
Bragança, Sofia M.
7df84b21-9316-4ee2-9445-4746df073fd8
Azevedo, Nuno F.
79da2c36-313d-41a4-84a1-caaa3f6b0264
Simöes, Lucia Chaves
9dd958fa-f504-4849-9975-fedfc3e9a013
Keevil, C. W.
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Vieira, M. J.
45deadf9-bf20-4e60-95e2-506226f0fed2

Bragança, Sofia M., Azevedo, Nuno F., Simöes, Lucia Chaves, Keevil, C. W. and Vieira, M. J. (2007) Use of fluorescent in situ hybridisation for the visualisation of Helicobacter pylori in real drinking water biofilms. Water Science & Technology, 55 (8-9), 387-393. (doi:10.2166/wst.2007.282).

Record type: Article

Abstract

A fluorescently labelled peptide nucleic acid (PNA) probe has been applied for the in situ detection of Helicobacter pylori in drinking water biofilms. The method was originally applied to real pipe samples removed from a drinking water distribution system (DWDS) but the curvature and the heavy fouling of the pipes prevented an accurate detection of the bacterium by epifluorescence microscopy. Therefore, two semi-circular flow cells were placed in a bypass of the DWDS, and coupons with up to 72 days of exposure were regularly sampled and analysed for the presence of H. pylori. In the flat surfaces of the coupons, it was possible to sparsely detect cells exhibiting similar morphology to H. pylori that were emitting the PNA probe fluorescent signal. Coupons were also visualised under the microscope before the hybridisation procedure to serve as negative controls and ensure the validity of the method. This work corroborates the findings already published elsewhere that this bacterium might be present in DWDS biofilms. The method requires, however, highly trained personnel for an accurate detection of the pathogen and will need simplification before being routinely used in standard water analysis laboratories. © IWA Publishing 2007.

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More information

Published date: 1 April 2007
Keywords: Biofilms, Drinking water, Helicobacter pylori, Peptide nucleic acids, Rapid detection methods

Identifiers

Local EPrints ID: 429847
URI: https://eprints.soton.ac.uk/id/eprint/429847
ISSN: 0273-1223
PURE UUID: 4900b095-a0c9-482a-8d18-627f4ea2bbbd
ORCID for C. W. Keevil: ORCID iD orcid.org/0000-0003-1917-7706

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Date deposited: 08 Apr 2019 16:30
Last modified: 09 Apr 2019 00:36

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Contributors

Author: Sofia M. Bragança
Author: Nuno F. Azevedo
Author: Lucia Chaves Simöes
Author: C. W. Keevil ORCID iD
Author: M. J. Vieira

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