The University of Southampton
University of Southampton Institutional Repository

PNA-FISH as a new diagnostic method for the determination of clarithromycin resistance of Helicobacter pylori

PNA-FISH as a new diagnostic method for the determination of clarithromycin resistance of Helicobacter pylori
PNA-FISH as a new diagnostic method for the determination of clarithromycin resistance of Helicobacter pylori
Background: triple therapy is the gold standard treatment for Helicobacter pylori eradication from the human stomach, but increased resistance to clarithromycin became the main factor of treatment failure. Until now, fastidious culturing methods are generally the method of choice to assess resistance status. In this study, a new genotypic method to detect clarithromycin resistance in clinical samples, based on fluorescent in situ hybridization (FISH) using a set of peptide nucleic acid probes (PNA), is proposed.

Results: the set of probes targeting the point mutations responsible for clarithromycin resistance was applied to H. pylori suspensions and showed 100% sensitivity and specificity (95% CI, 79.9-100 and 95% CI, 71.6-100 respectively). This method can also be amenable for application to gastric biopsy samples, as resistance to clarithromycin was also detected when histological slides were tested.

Conclusions: the optimized PNA-FISH based diagnostic method to detect H. pylori clarithromycin resistance shown to be a very sensitive and specific method for the detection of clarithromycin resistance in the H. pylori smears and also proved to be a reliable method for the diagnosis of this pathogen in clinical samples and an alternative to existing plating methods
101-[7pp]
Cerqueira, Laura
61343b41-4742-4057-94e9-2ef4ea145613
Fernandes, Ricardo M..
d9c3de62-729d-44d6-8616-47f5096fd7c2
Ferreira, Rui M.
8ec38033-099b-43e3-80b9-2c736b6e3394
Carneiro, Fátima
09da70a4-8ca4-4078-b177-f3ccd870b447
Dinis-Ribeiro, Mário
7d966cbc-870e-43cd-a3c4-6685599d27bd
Figueiredo, Céu
54d5e121-a8e1-48f1-93d0-453b7fa9a97f
Keevil, Charles W.
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Azevedo, Nuno F.
24c4eb52-0c98-443b-881f-7a1449c9ac26
Vieira, Maria J.
d972e877-d85b-488c-8b0f-358f79d2fa29
Cerqueira, Laura
61343b41-4742-4057-94e9-2ef4ea145613
Fernandes, Ricardo M..
d9c3de62-729d-44d6-8616-47f5096fd7c2
Ferreira, Rui M.
8ec38033-099b-43e3-80b9-2c736b6e3394
Carneiro, Fátima
09da70a4-8ca4-4078-b177-f3ccd870b447
Dinis-Ribeiro, Mário
7d966cbc-870e-43cd-a3c4-6685599d27bd
Figueiredo, Céu
54d5e121-a8e1-48f1-93d0-453b7fa9a97f
Keevil, Charles W.
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Azevedo, Nuno F.
24c4eb52-0c98-443b-881f-7a1449c9ac26
Vieira, Maria J.
d972e877-d85b-488c-8b0f-358f79d2fa29

Cerqueira, Laura, Fernandes, Ricardo M.., Ferreira, Rui M., Carneiro, Fátima, Dinis-Ribeiro, Mário, Figueiredo, Céu, Keevil, Charles W., Azevedo, Nuno F. and Vieira, Maria J. (2011) PNA-FISH as a new diagnostic method for the determination of clarithromycin resistance of Helicobacter pylori. BMC Microbiology, 11 (1), 101-[7pp]. (doi:10.1186/1471-2180-11-101).

Record type: Article

Abstract

Background: triple therapy is the gold standard treatment for Helicobacter pylori eradication from the human stomach, but increased resistance to clarithromycin became the main factor of treatment failure. Until now, fastidious culturing methods are generally the method of choice to assess resistance status. In this study, a new genotypic method to detect clarithromycin resistance in clinical samples, based on fluorescent in situ hybridization (FISH) using a set of peptide nucleic acid probes (PNA), is proposed.

Results: the set of probes targeting the point mutations responsible for clarithromycin resistance was applied to H. pylori suspensions and showed 100% sensitivity and specificity (95% CI, 79.9-100 and 95% CI, 71.6-100 respectively). This method can also be amenable for application to gastric biopsy samples, as resistance to clarithromycin was also detected when histological slides were tested.

Conclusions: the optimized PNA-FISH based diagnostic method to detect H. pylori clarithromycin resistance shown to be a very sensitive and specific method for the detection of clarithromycin resistance in the H. pylori smears and also proved to be a reliable method for the diagnosis of this pathogen in clinical samples and an alternative to existing plating methods

PDF
1471-2180-11-101.pdf - Version of Record
Available under License Other.
Download (1MB)

More information

Published date: 14 May 2011
Organisations: Centre for Biological Sciences

Identifiers

Local EPrints ID: 209291
URI: https://eprints.soton.ac.uk/id/eprint/209291
PURE UUID: f16928eb-bc65-4b72-a0a0-893a4cc3b73a
ORCID for Charles W. Keevil: ORCID iD orcid.org/0000-0003-1917-7706

Catalogue record

Date deposited: 27 Jan 2012 11:35
Last modified: 06 Jun 2018 12:49

Export record

Altmetrics

Contributors

Author: Laura Cerqueira
Author: Ricardo M.. Fernandes
Author: Rui M. Ferreira
Author: Fátima Carneiro
Author: Mário Dinis-Ribeiro
Author: Céu Figueiredo
Author: Nuno F. Azevedo
Author: Maria J. Vieira

University divisions

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of https://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×