Fluorescence In Vivo Hybridization (FIVH) for Detection of Helicobacter pylori Infection in a C57BL/6 Mouse Model

• Type: Article in International Scientific Journal
• Date: 2016

Authors

Sílvia Fontenete (Author) FEUP; Marina Leite (Author) Other; Davie Cappoen (Author) Other; Rita Santos (Author) FEUP; Chris Van Ginneken (Author) Other; Céu Figueiredo (Author) FMUP; Jesper Wengel (Author) Other; Paul Cos (Author) Other; Nuno Filipe Azevedo (Author) FEUP

Journal

Title: PLoS ONE

Serial No. e0145669 Vol. 11 No. 2

Pages: 1-18

ISSN: 1932-6203

Publisher: Public Library of Science

Indexing

ISI Web of Knowledge - 12 Citations

Scopus - 13 Citations

Associated Projects

LEPAE/CEFT - RL2 - (Bio) Chemical Engineering: Multi-Scale Approaches for Sustainable Environment and Health

DNA mimics - Desenvolvimento e aplicação de mímicos de DNA para a rápida identificação de microrganismos patogénicos

LEPABE - Laboratório de Engenharia de Processos, Ambiente, Biotecnologia e Energia

Other information

Authenticus ID: P-00K-5HZ

DOI: 10.1371/journal.pone.0148353

Abstract (EN): Introduction In this study, we applied fluorescence in vivo hybridization (FIVH) using locked nucleic acid (LNA) probes targeting the bacterial rRNA gene for in vivo detection of H. pylori infecting the C57BL/6 mouse model. A previously designed Cy3_HP_LNA/2OMe_PS probe, complementary to a sequence of the H. pylori 16S rRNA gene, was used. First, the potential cytotoxicity and genotoxicity of the probe was assessed by commercial assays. Further, the performance of the probe for detecting H. pylori at different pH conditions was tested in vitro, using fluorescence in situ hybridization (FISH). Finally, the efficiency of FIVH to detect H. pylori SS1 strain in C57BL/6 infectedmice was evaluated ex vivo in mucus samples, in cryosections and paraffin-embedded sections by epifluorescence and confocal microscopy. Results H. pylori SS1 strain infecting C57BL/6 mice was successfully detected by the Cy3_HP_LNA/2OMe_PS probe in the mucus, attached to gastric epithelial cells and colonizing the gastric pits. The specificity of the probe for H. pylori was confirmed by microscopy. Conclusions In the future this methodology can be used in combination with a confocal laser endomicroscope for in vivo diagnosis of H. pylori infection using fluorescent LNA probes, which would be helpful to obtain an immediate diagnosis. Our results proved for the first time that FIVH method is applicable inside the body of a higher-order animal.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 18