title: Fluorescence In Vivo Hybridization (FIVH) for Detection of Helicobacter pylori Infection in a C57BL/6 Mouse Model.
abstract: BACKGROUND
  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 infected mice 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.
authors: LEPABE, Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Porto, Portugal.|i3S, Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|IPATIMUP, Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal.|Nucleic Acid Center, Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Odense M, Denmark.|ICBAS, Institute of Biomedical Sciences Abel Salazar, University of Porto, Porto, Portugal.
authors: Sílvia
authors: S
authors: Fontenete
authors: i3S, Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|IPATIMUP, Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal.
authors: Marina
authors: M
authors: Leite
authors: Laboratory of Microbiology, Parasitology and Hygiene (LMPH), Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, University of Antwerp, Antwerp, Belgium.
authors: Davie
authors: D
authors: Cappoen
authors: LEPABE, Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Porto, Portugal.|i3S, Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|IPATIMUP, Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal.|Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Gent, Belgium.
authors: Rita
authors: R
authors: Santos
authors: Laboratory of Applied Veterinary Morphology, Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, University of Antwerp, Antwerp, Belgium.
authors: Chris Van
authors: CV
authors: Ginneken
authors: i3S, Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|IPATIMUP, Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal.|FMUP, Faculty of Medicine of the University of Porto, University, Porto, Portugal.
authors: Céu
authors: C
authors: Figueiredo
authors: Nucleic Acid Center, Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Odense M, Denmark.
authors: Jesper
authors: J
authors: Wengel
authors: Laboratory of Microbiology, Parasitology and Hygiene (LMPH), Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, University of Antwerp, Antwerp, Belgium.
authors: Paul
authors: P
authors: Cos
authors: LEPABE, Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Porto, Portugal.
authors: Nuno Filipe
authors: NF
authors: Azevedo
mesh_terms: D000818:Animals; D002460:Cell Line; D004195:Disease Models, Animal; D005260:Female; D005753:Gastric Mucosa / Q000382:microbiology; D016481:Helicobacter Infections / Q000175:diagnosis*; D016480:Helicobacter pylori / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D009841:Oligonucleotides / Q000378:metabolism; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D009841:Oligonucleotides; D012329:RNA, Bacterial; C477371:locked nucleic acid