Towards Fluorescence In Vivo Hybridization (FIVH) Detection of H. pylori in Gastric Mucosa Using Advanced LNA Probes
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{"title"=>"Towards fluorescence in vivo hybridization (FIVH) detection of H. pylori in gastric mucosa using advanced LNA probes", "type"=>"journal", "authors"=>[{"first_name"=>"Sílvia", "last_name"=>"Fontenete", "scopus_author_id"=>"55216563200"}, {"first_name"=>"Marina", "last_name"=>"Leite", "scopus_author_id"=>"56436933000"}, {"first_name"=>"Nuno", "last_name"=>"Guimarães", "scopus_author_id"=>"17345683500"}, {"first_name"=>"Pedro", "last_name"=>"Madureira", "scopus_author_id"=>"6506395469"}, {"first_name"=>"Rui Manuel", "last_name"=>"Ferreira", "scopus_author_id"=>"34976443500"}, {"first_name"=>"Céu", "last_name"=>"Figueiredo", "scopus_author_id"=>"7006520927"}, {"first_name"=>"Jesper", "last_name"=>"Wengel", "scopus_author_id"=>"7103094424"}, {"first_name"=>"Nuno Filipe", "last_name"=>"Azevedo", "scopus_author_id"=>"8092231700"}], "year"=>2015, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"604006411", "issn"=>"19326203", "pmid"=>"2015983842", "sgr"=>"84928573017", "scopus"=>"2-s2.0-84928573017", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0125494"}, "id"=>"ba468d4e-e9d0-3904-945e-47549fdb3b8b", "abstract"=>"In recent years, there have been several attempts to improve the diagnosis of infection caused by Helicobacter pylori. Fluorescence in situ hybridization (FISH) is a commonly used technique to detect H. pylori infection but it requires biopsies from the stomach. Thus, the development of an in vivo FISH-based method (FIVH) that directly detects and allows the visualization of the bacterium within the human body would significantly reduce the time of analysis, allowing the diagnosis to be performed during endoscopy. In a previous study we designed and synthesized a phosphorothioate locked nucleic acid (LNA)/ 2′ O-methyl RNA (2′OMe) probe using standard phosphoramidite chemistry and FISH hybridization was then successfully performed both on adhered and suspended bacteria at 37°C. In this work we simplified, shortened and adapted FISH to work at gastric pH values, meaning that the hybridization step now takes only 30 minutes and, in addition to the buffer, uses only urea and probe at non-toxic concentrations. Importantly, the sensitivity and specificity of the FISH method was maintained in the range of conditions tested, even at low stringency conditions (e.g., low pH). In conclusion, this methodology is a promising approach that might be used in vivo in the future in combination with a confocal laser endomicroscope for H. pylori visualization. © 2015 Fontenete et al.", "link"=>"http://www.mendeley.com/research/towards-fluorescence-vivo-hybridization-fivh-detection-h-pylori-gastric-mucosa-using-advanced-lna-pr", "reader_count"=>17, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>3, "Student > Master"=>6, "Student > Bachelor"=>1, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>3, "Student > Master"=>6, "Student > Bachelor"=>1, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>6, "Agricultural and Biological Sciences"=>5, "Medicine and Dentistry"=>1, "Earth and Planetary Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Earth and Planetary Sciences"=>{"Earth and Planetary Sciences"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>5}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}, "Unspecified"=>{"Unspecified"=>3}}, "reader_count_by_country"=>{"United States"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/2040708"], "description"=>"<p>Three dimensional surface plot showing the variation of <i>H</i>. <i>pylori</i> fluorescence intensity as a function of model terms. The model graphs are represented in gradient color shading. The surface is red at higher response levels and blue at lower ones.</p>", "links"=>[], "tags"=>["FISH hybridization", "probe", "Advanced LNA Probes", "confocal laser endomicroscope", "hybridization step", "Gastric mucosa", "FIVH", "Towards Fluorescence", "rna", "pylori visualization", "Vivo Hybridization", "vivo", "pylori infection", "Helicobacter pylori", "stringency conditions", "FISH method", "pH values", "phosphoramidite chemistry", "30 minutes", "diagnosis"], "article_id"=>1395130, "categories"=>["Biological Sciences"], "users"=>["Sílvia Fontenete", "Marina Leite", "Nuno Guimarães", "Pedro Madureira", "Rui Manuel Ferreira", "Ceu Figueiredo", "Jesper Wengel", "Nuno Filipe Azevedo"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0125494.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_FAM_HP_LNA_2OMe_PS_probe_performance_using_the_response_surface_methodology_RSM_/1395130", "title"=>"Analysis of FAM HP_LNA/2OMe_PS probe performance using the response surface methodology (RSM).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-04-27 02:48:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/2040709"], "description"=>"<p>A—F Smear of pure culture of <i>H</i>. <i>pylori</i> strain 26695 observed by epifluorescent microscopy. A-C. Experiment using 200 nM of the probe. D-F. Smears without probe were used as negative control. All images were taken at equal exposure times. G-I. Relative fluorescence histograms of LNA-FISH targeting <i>H</i>. <i>pylori</i> in different pH for two different assays—Blue: negative control with no probe; Green: positive sample. Scale bar = 10 μm.</p>", "links"=>[], "tags"=>["FISH hybridization", "probe", "Advanced LNA Probes", "confocal laser endomicroscope", "hybridization step", "Gastric mucosa", "FIVH", "Towards Fluorescence", "rna", "pylori visualization", "Vivo Hybridization", "vivo", "pylori infection", "Helicobacter pylori", "stringency conditions", "FISH method", "pH values", "phosphoramidite chemistry", "30 minutes", "diagnosis"], "article_id"=>1395131, "categories"=>["Biological Sciences"], "users"=>["Sílvia Fontenete", "Marina Leite", "Nuno Guimarães", "Pedro Madureira", "Rui Manuel Ferreira", "Ceu Figueiredo", "Jesper Wengel", "Nuno Filipe Azevedo"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0125494.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Detection_of_H_pylori_in_slides_by_epifluorescent_microscopy_A_F_and_in_suspension_by_flow_cytometry_H_J_using_the_FAM_HP_LNA_2OMe_PS_oligonucleotides_probe_at_different_pH_values_/1395131", "title"=>"Detection of <i>H</i>. <i>pylori</i> in slides, by epifluorescent microscopy (A-F), and in suspension, by flow cytometry (H-J), using the FAM HP_ LNA/2OMe _PS oligonucleotides probe at different pH values.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-04-27 02:48:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/2040710"], "description"=>"<p>A. Sensitivity test using different strains and clinical isolates of <i>H</i>. <i>pylori</i>. C. Analysis of the probe in clinical isolates of <i>H</i>. <i>pylori</i>. B. Specificity test using different species of Helicobacter and other bacteria.</p>", "links"=>[], "tags"=>["FISH hybridization", "probe", "Advanced LNA Probes", "confocal laser endomicroscope", "hybridization step", "Gastric mucosa", "FIVH", "Towards Fluorescence", "rna", "pylori visualization", "Vivo Hybridization", "vivo", "pylori infection", "Helicobacter pylori", "stringency conditions", "FISH method", "pH values", "phosphoramidite chemistry", "30 minutes", "diagnosis"], "article_id"=>1395132, "categories"=>["Biological Sciences"], "users"=>["Sílvia Fontenete", "Marina Leite", "Nuno Guimarães", "Pedro Madureira", "Rui Manuel Ferreira", "Ceu Figueiredo", "Jesper Wengel", "Nuno Filipe Azevedo"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0125494.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Sensitivity_and_specificity_studies_performed_with_the_HP_LNA_2OMe_PS_probe_at_different_pH_values_evaluated_by_flow_cytometry_/1395132", "title"=>"Sensitivity and specificity studies performed with the HP_ LNA/2OMe _PS probe at different pH values evaluated by flow cytometry.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-04-27 02:48:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/2040711"], "description"=>"<p>A-F Smear of pure culture of <i>H</i>. <i>pylori</i> strain 26695 (ATCC 700392) observed by epifluorescent microscopy. A-C Experiment using 200 nM of Cy3 HP_ LNA/2OMe _PS oligonucleotide probe. D-F. Smears without probe were used as negative control. All images were taken at equal exposure times. Original magnification: 1000x. G-I. Relative fluorescence histograms of LNA-FISH targeting <i>H</i>. <i>pylori</i> in different pH for two different assays—Blue: negative control with no oligonucleotide probe; Red: sample. Scale bar = 5 μm.</p>", "links"=>[], "tags"=>["FISH hybridization", "probe", "Advanced LNA Probes", "confocal laser endomicroscope", "hybridization step", "Gastric mucosa", "FIVH", "Towards Fluorescence", "rna", "pylori visualization", "Vivo Hybridization", "vivo", "pylori infection", "Helicobacter pylori", "stringency conditions", "FISH method", "pH values", "phosphoramidite chemistry", "30 minutes", "diagnosis"], "article_id"=>1395133, "categories"=>["Biological Sciences"], "users"=>["Sílvia Fontenete", "Marina Leite", "Nuno Guimarães", "Pedro Madureira", "Rui Manuel Ferreira", "Ceu Figueiredo", "Jesper Wengel", "Nuno Filipe Azevedo"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0125494.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Detection_of_H_pylori_using_the_Cy3_HP_LNA_2OMe_PS_oligonucleotide_probe_in_a_smears_of_pure_culture_of_H_pylori_strain_26695_using_simulated_gastric_juice_by_epifluorescent_microscopy_/1395133", "title"=>"Detection of <i>H</i>. <i>pylori</i> using the Cy3 HP_ LNA/2OMe _PS oligonucleotide probe in a smears of pure culture of <i>H</i>. <i>pylori</i> strain 26695 using simulated gastric juice by epifluorescent microscopy.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-04-27 02:48:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/2040712"], "description"=>"<p>A. Sensitivity test using different strains and clinical isolates of <i>H</i>. <i>pylori</i>. B. Specificity test using different species of Helicobacter and other bacteria.</p>", "links"=>[], "tags"=>["FISH hybridization", "probe", "Advanced LNA Probes", "confocal laser endomicroscope", "hybridization step", "Gastric mucosa", "FIVH", "Towards Fluorescence", "rna", "pylori visualization", "Vivo Hybridization", "vivo", "pylori infection", "Helicobacter pylori", "stringency conditions", "FISH method", "pH values", "phosphoramidite chemistry", "30 minutes", "diagnosis"], "article_id"=>1395134, "categories"=>["Biological Sciences"], "users"=>["Sílvia Fontenete", "Marina Leite", "Nuno Guimarães", "Pedro Madureira", "Rui Manuel Ferreira", "Ceu Figueiredo", "Jesper Wengel", "Nuno Filipe Azevedo"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0125494.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Sensitivity_and_specificity_studies_performed_with_the_HP_LNA_2OMe_PS_oligonucleotide_probe_using_simulated_gastric_juice_/1395134", "title"=>"Sensitivity and specificity studies performed with the HP_ LNA/2OMe _PS oligonucleotide probe using simulated gastric juice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-04-27 02:48:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/2040713"], "description"=>"<p>A. Detection of <i>H</i>. <i>pylori</i> by the Cy3_ HP_ LNA/2OMe _PS _oligonucleotide probe in infected epithelial cells. Channel red, DAPI and bright field were overlapping. Scale bar = 10μm B. Detection of <i>H</i>. <i>pylori</i> by Cy3_ HP_ LNA/2OMe _PS _oligonucleotide probe in co-infected cells and isolates bacteria. Channel red, and bright field were overlapping. Scale bar = 25μm C. Uninfected AGS cells stained with Cy3_ HP_ LNA/2OMe _PS _oligonucleotide probe and DAPI. Channel red, and bright field and DAPI were overlapping. Scale bar = 5μm. Red: Cy3 fluorescence. Blue: DAPI staining to counterstain nuclei.</p>", "links"=>[], "tags"=>["FISH hybridization", "probe", "Advanced LNA Probes", "confocal laser endomicroscope", "hybridization step", "Gastric mucosa", "FIVH", "Towards Fluorescence", "rna", "pylori visualization", "Vivo Hybridization", "vivo", "pylori infection", "Helicobacter pylori", "stringency conditions", "FISH method", "pH values", "phosphoramidite chemistry", "30 minutes", "diagnosis"], "article_id"=>1395135, "categories"=>["Biological Sciences"], "users"=>["Sílvia Fontenete", "Marina Leite", "Nuno Guimarães", "Pedro Madureira", "Rui Manuel Ferreira", "Ceu Figueiredo", "Jesper Wengel", "Nuno Filipe Azevedo"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0125494.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Confocal_microscopy_images_of_AGS_cells_infected_with_H_pylori_26695_strain_/1395135", "title"=>"Confocal microscopy images of AGS cells infected with <i>H</i>. <i>pylori</i> 26695 strain.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-04-27 02:48:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/2040714"], "description"=>"<p>AGS cells were treated with 200 nM of Cy3_ HP_ LNA/2OMe _PS oligonucleotide probe for 24h. The results are the mean ± SEM of three independent experiments; *<i>p<0</i>.<i>05 vs</i> untreated cells by ANOVA. A) Cell viability was measured using MTS assay. B) Analysis of caspase-3/7 activity in AGS cells treated with Cy3_ HP_ LNA/2OMe _PS oligonucleotide probe. Staurosporine, STS, was a positive control of cell death. C) Effect of Cy3_ HP_ LNA/2OMe _PS oligonucleotide probe on the amount of DNA fragmentation in the cultured gastric cell line AGS. The level of apoptosis occurring with each treatment was determined by cell death ELISA<sup>Plus</sup> kit.</p>", "links"=>[], "tags"=>["FISH hybridization", "probe", "Advanced LNA Probes", "confocal laser endomicroscope", "hybridization step", "Gastric mucosa", "FIVH", "Towards Fluorescence", "rna", "pylori visualization", "Vivo Hybridization", "vivo", "pylori infection", "Helicobacter pylori", "stringency conditions", "FISH method", "pH values", "phosphoramidite chemistry", "30 minutes", "diagnosis"], "article_id"=>1395136, "categories"=>["Biological Sciences"], "users"=>["Sílvia Fontenete", "Marina Leite", "Nuno Guimarães", "Pedro Madureira", "Rui Manuel Ferreira", "Ceu Figueiredo", "Jesper Wengel", "Nuno Filipe Azevedo"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0125494.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_the_Cy3_HP_LNA_2OMe_PS_oligonucleotide_probe_on_viability_MTS_assay_A_and_cell_death_apoptosis_B_and_C_of_AGS_cells_/1395136", "title"=>"Effect of the Cy3_ HP_ LNA/2OMe _PS oligonucleotide probe on viability (MTS assay, A) and cell death (apoptosis, B and C) of AGS cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-04-27 02:48:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/2040715"], "description"=>"<p>HP_ LNA/2OMe _PS is a phosphorothioate oligomer (PS backbones) labeled with either FAM (Fluorescein) or Cy3 (Cyanine).</p><p>Designation and sequence of the oligonucleotide probes containing locked nucleic acid (LNA; with L superscript) and 2’-O-methyl RNA (2’-OMe; in Boldface) nucleotide monomers.</p>", "links"=>[], "tags"=>["FISH hybridization", "probe", "Advanced LNA Probes", "confocal laser endomicroscope", "hybridization step", "Gastric mucosa", "FIVH", "Towards Fluorescence", "rna", "pylori visualization", "Vivo Hybridization", "vivo", "pylori infection", "Helicobacter pylori", "stringency conditions", "FISH method", "pH values", "phosphoramidite chemistry", "30 minutes", "diagnosis"], "article_id"=>1395137, "categories"=>["Biological Sciences"], "users"=>["Sílvia Fontenete", "Marina Leite", "Nuno Guimarães", "Pedro Madureira", "Rui Manuel Ferreira", "Ceu Figueiredo", "Jesper Wengel", "Nuno Filipe Azevedo"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0125494.t001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Designation_and_sequence_of_the_oligonucleotide_probes_containing_locked_nucleic_acid_LNA_with_L_superscript_and_2_8217_O_methyl_RNA_2_8217_OMe_in_Boldface_nucleotide_monomers_/1395137", "title"=>"Designation and sequence of the oligonucleotide probes containing locked nucleic acid (LNA; with L superscript) and 2’-O-methyl RNA (2’-OMe; in Boldface) nucleotide monomers.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2015-04-27 02:48:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/2040716"], "description"=>"<p>Legend: <i>Helicobacter</i> clinical isolates provided by:</p><p><sup>a</sup> Céu Figueiredo [<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125494#pone.0125494.ref028\" target=\"_blank\">28</a>];</p><p><sup>b</sup> Francis Megraud, and</p><p><sup>c</sup> Jay Solnick</p><p><i>Helicobacter</i> and non-<i>Helicobacter</i> bacterial strains included in this study.</p>", "links"=>[], "tags"=>["FISH hybridization", "probe", "Advanced LNA Probes", "confocal laser endomicroscope", "hybridization step", "Gastric mucosa", "FIVH", "Towards Fluorescence", "rna", "pylori visualization", "Vivo Hybridization", "vivo", "pylori infection", "Helicobacter pylori", "stringency conditions", "FISH method", "pH values", "phosphoramidite chemistry", "30 minutes", "diagnosis"], "article_id"=>1395138, "categories"=>["Biological Sciences"], "users"=>["Sílvia Fontenete", "Marina Leite", "Nuno Guimarães", "Pedro Madureira", "Rui Manuel Ferreira", "Ceu Figueiredo", "Jesper Wengel", "Nuno Filipe Azevedo"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0125494.t002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Helicobacter_and_non_Helicobacter_bacterial_strains_included_in_this_study_/1395138", "title"=>"<i>Helicobacter</i> and non-<i>Helicobacter</i> bacterial strains included in this study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2015-04-27 02:48:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/2040717", "https://ndownloader.figshare.com/files/2040718", "https://ndownloader.figshare.com/files/2040719", "https://ndownloader.figshare.com/files/2040720", "https://ndownloader.figshare.com/files/2040721", "https://ndownloader.figshare.com/files/2040722", "https://ndownloader.figshare.com/files/2040723"], "description"=>"<div><p>In recent years, there have been several attempts to improve the diagnosis of infection caused by <i>Helicobacter pylori</i>. Fluorescence <i>in situ</i> hybridization (FISH) is a commonly used technique to detect <i>H</i>. <i>pylori</i> infection but it requires biopsies from the stomach. Thus, the development of an <i>in vivo</i> FISH-based method (FIVH) that directly detects and allows the visualization of the bacterium within the human body would significantly reduce the time of analysis, allowing the diagnosis to be performed during endoscopy. In a previous study we designed and synthesized a phosphorothioate locked nucleic acid (LNA)/ 2’ O-methyl RNA (2’OMe) probe using standard phosphoramidite chemistry and FISH hybridization was then successfully performed both on adhered and suspended bacteria at 37°C. In this work we simplified, shortened and adapted FISH to work at gastric pH values, meaning that the hybridization step now takes only 30 minutes and, in addition to the buffer, uses only urea and probe at non-toxic concentrations. Importantly, the sensitivity and specificity of the FISH method was maintained in the range of conditions tested, even at low stringency conditions (e.g., low pH). In conclusion, this methodology is a promising approach that might be used <i>in vivo</i> in the future in combination with a confocal laser endomicroscope for <i>H</i>. <i>pylori</i> visualization.</p></div>", "links"=>[], "tags"=>["FISH hybridization", "probe", "Advanced LNA Probes", "confocal laser endomicroscope", "hybridization step", "Gastric mucosa", "FIVH", "Towards Fluorescence", "rna", "pylori visualization", "Vivo Hybridization", "vivo", "pylori infection", "Helicobacter pylori", "stringency conditions", "FISH method", "pH values", "phosphoramidite chemistry", "30 minutes", "diagnosis"], "article_id"=>1395139, "categories"=>["Biological Sciences"], "users"=>["Sílvia Fontenete", "Marina Leite", "Nuno Guimarães", "Pedro Madureira", "Rui Manuel Ferreira", "Ceu Figueiredo", "Jesper Wengel", "Nuno Filipe Azevedo"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0125494.s001", "https://dx.doi.org/10.1371/journal.pone.0125494.s002", "https://dx.doi.org/10.1371/journal.pone.0125494.s003", "https://dx.doi.org/10.1371/journal.pone.0125494.s004", "https://dx.doi.org/10.1371/journal.pone.0125494.s005", "https://dx.doi.org/10.1371/journal.pone.0125494.s006", "https://dx.doi.org/10.1371/journal.pone.0125494.s007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Towards_Fluorescence_In_Vivo_Hybridization_FIVH_Detection_of_H_pylori_in_Gastric_Mucosa_Using_Advanced_LNA_Probes/1395139", "title"=>"Towards Fluorescence <i>In Vivo</i> Hybridization (FIVH) Detection of <i>H</i>. <i>pylori</i> in Gastric Mucosa Using Advanced LNA Probes", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-04-27 02:48:24"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"16", "full-text"=>"17", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"12"}
  • {"unique-ip"=>"9", "full-text"=>"10", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"9", "full-text"=>"10", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"18", "full-text"=>"18", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"12", "full-text"=>"12", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"13", "full-text"=>"11", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"12", "full-text"=>"11", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"7", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}

Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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