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

BMC Microbiology

Methodology Article

Charles W Keevil¹  Laura Cerqueira²  Maria J Vieira²  Ricardo M Fernandes²  Nuno F Azevedo³  Céu Figueiredo⁴  Rui M Ferreira⁴  Fátima Carneiro⁵  Mário Dinis-Ribeiro⁶ 

[1] Environmental Healthcare Unit, School of Biological Sciences, University of Southampton, Southampton, UK;IBB - Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar 4710-057, Braga, Portugal;IBB - Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar 4710-057, Braga, Portugal;LEPAE, Department of Chemical Engineering, Faculty of Engineering, University of Porto, Porto, Portugal;IPATIMUP - Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal;Medical Faculty of the University of Porto, Porto, Portugal;IPATIMUP - Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal;Medical Faculty of the University of Porto, Porto, Portugal;Department of Pathology, Hospital S. João, Porto, Portugal;Medical Faculty of the University of Porto, Porto, Portugal;Portuguese Oncology Institute Porto, Department Gastroenterology, Porto, Portugal;

关键词: Clarithromycin;    Pylorus Eradication;    Peptide Nucleic Acid;    Pylorus Strain;    Gastric Biopsy;   

DOI  :  10.1186/1471-2180-11-101

 received in 2011-02-23, accepted in 2011-05-14,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundTriple 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.ResultsThe 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.ConclusionsThe 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.

【 授权许可】

Unknown   
© Cerqueira et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.