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

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).


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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

Item Type: Article
Digital Object Identifier (DOI): doi:10.1186/1471-2180-11-101
ISSNs: 1471-2180 (print)
Subjects: Q Science > QR Microbiology
Divisions : Faculty of Natural and Environmental Sciences > Biological Sciences
ePrint ID: 209291
Accepted Date and Publication Date:
14 May 2011Published
Date Deposited: 27 Jan 2012 11:35
Last Modified: 31 Mar 2016 13:49

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