Detection of Prion Protein Particles in Blood Plasma of Scrapie Infected Sheep
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{"title"=>"Detection of prion protein particles in blood plasma of scrapie infected sheep", "type"=>"journal", "authors"=>[{"first_name"=>"Oliver", "last_name"=>"Bannach", "scopus_author_id"=>"14419945200"}, {"first_name"=>"Eva", "last_name"=>"Birkmann", "scopus_author_id"=>"24166196300"}, {"first_name"=>"Elke", "last_name"=>"Reinartz", "scopus_author_id"=>"55203010100"}, {"first_name"=>"Karl Erich", "last_name"=>"Jaeger", "scopus_author_id"=>"7103288943"}, {"first_name"=>"Jan P.M.", "last_name"=>"Langeveld", "scopus_author_id"=>"7006720028"}, {"first_name"=>"Robert G.", "last_name"=>"Rohwer", "scopus_author_id"=>"7003874958"}, {"first_name"=>"Luisa", "last_name"=>"Gregori", "scopus_author_id"=>"6701453414"}, {"first_name"=>"Linda A.", "last_name"=>"Terry", "scopus_author_id"=>"36959293700"}, {"first_name"=>"Dieter", "last_name"=>"Willbold", "scopus_author_id"=>"7004563027"}, {"first_name"=>"Detlev", "last_name"=>"Riesner", "scopus_author_id"=>"7005142635"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84860456864", "sgr"=>"84860456864", "pui"=>"364721418", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"22567169", "doi"=>"10.1371/journal.pone.0036620"}, "id"=>"24e62d21-9ec9-3ed5-b00e-d301362e0ecd", "abstract"=>"Prion diseases are transmissible neurodegenerative diseases affecting humans and animals. The agent of the disease is the prion consisting mainly, if not solely, of a misfolded and aggregated isoform of the host-encoded prion protein (PrP). Transmission of prions can occur naturally but also accidentally, e.g. by blood transfusion, which has raised serious concerns about blood product safety and emphasized the need for a reliable diagnostic test. In this report we present a method based on surface-FIDA (fluorescence intensity distribution analysis), that exploits the high state of molecular aggregation of PrP as an unequivocal diagnostic marker of the disease, and show that it can detect infection in blood. To prepare PrP aggregates from blood plasma we introduced a detergent and lipase treatment to separate PrP from blood lipophilic components. Prion protein aggregates were subsequently precipitated by phosphotungstic acid, immobilized on a glass surface by covalently bound capture antibodies, and finally labeled with fluorescent antibody probes. Individual PrP aggregates were visualized by laser scanning microscopy where signal intensity was proportional to aggregate size. After signal processing to remove the background from low fluorescence particles, fluorescence intensities of all remaining PrP particles were summed. We detected PrP aggregates in plasma samples from six out of ten scrapie-positive sheep with no false positives from uninfected sheep. Applying simultaneous intensity and size discrimination, ten out of ten samples from scrapie sheep could be differentiated from uninfected sheep. The implications for ante mortem diagnosis of prion diseases are discussed.", "link"=>"http://www.mendeley.com/research/detection-prion-protein-particles-blood-plasma-scrapie-infected-sheep", "reader_count"=>33, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Researcher"=>11, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>6, "Student > Master"=>6, "Other"=>3, "Student > Bachelor"=>3, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Researcher"=>11, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>6, "Student > Master"=>6, "Other"=>3, "Student > Bachelor"=>3, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>17, "Medicine and Dentistry"=>1, "Neuroscience"=>1, "Veterinary Science and Veterinary Medicine"=>2, "Physics and Astronomy"=>1, "Immunology and Microbiology"=>1, "Earth and Planetary Sciences"=>1, "Economics, Econometrics and Finance"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Neuroscience"=>{"Neuroscience"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Earth and Planetary Sciences"=>{"Earth and Planetary Sciences"=>1}, "Economics, Econometrics and Finance"=>{"Economics, Econometrics and Finance"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>17}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Unspecified"=>{"Unspecified"=>3}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>2}}, "reader_count_by_country"=>{"United States"=>1, "Italy"=>2}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/643643"], "description"=>"<p>Replicate samples of a plasma pool prepared from sheep symptomatic for scrapie and from a pool prepared from uninfected control sheep were processed independently by two experimenters (A and B). Discriminability of positive and negative samples is expressed as fluorescence intensity (positive, p) minus intensity (negative, n) divided by intensity (positive, p). MAb SAF-32 was used as capture, mAb L42 as probe.</p>", "links"=>[], "tags"=>["plasma"], "article_id"=>314128, "categories"=>["Biochemistry", "Infectious Diseases"], "users"=>["Oliver Bannach", "Eva Birkmann", "Elke Reinartz", "Karl-Erich Jaeger", "Jan P. M. Langeveld", "Robert G. Rohwer", "Luisa Gregori", "Linda A. Terry", "Dieter Willbold", "Detlev Riesner"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036620.g003", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Reproducibility_of_plasma_analyses_/314128", "title"=>"Reproducibility of plasma analyses.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:38:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/643532"], "description"=>"<p>To better resolve grayscales, images are colorized with ‘fire’ lookup table (cf. right bar). Each sample contained 50 ng recPrP aggregates in 100 µl sample volume. All samples were analyzed by surface-FIDA assay using SAF-32 as capture and detection antibody. One image corresponds to 1% of the total well bottom area. RecPrP aggregates (<b>A</b>) in PBS, (<b>B</b>) in ovine plasma, (<b>C</b>) in plasma and 2% sarkosyl, (<b>D</b>) in plasma after sarkosyl and lipase treatment, (<b>E</b>) in plasma after sarkosyl and lipase treatment and PTA-precipitation, (<b>F</b>) in plasma after PTA precipitation. (<b>G</b>) negative control: non-spiked plasma after sarkosyl, lipase and PTA steps.</p>", "links"=>[], "tags"=>["recprp", "particles"], "article_id"=>314017, "categories"=>["Biochemistry", "Infectious Diseases"], "users"=>["Oliver Bannach", "Eva Birkmann", "Elke Reinartz", "Karl-Erich Jaeger", "Jan P. M. Langeveld", "Robert G. Rohwer", "Luisa Gregori", "Linda A. Terry", "Dieter Willbold", "Detlev Riesner"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036620.g002", "stats"=>{"downloads"=>2, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Detection_of_recPrP_particles_in_the_presence_of_blood_plasma_/314017", "title"=>"Detection of recPrP particles in the presence of blood plasma.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:37:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/643426"], "description"=>"<p>A ‘rolling ball’ moves along (arrow) the underside of the intensity curve and thereby identifies the background, which is subsequently subtracted. Remaining peaks of low intensities are removed by an intensity cut-off (dashed line). In the algorithm, the ball moves along a three-dimensional landscape, identifying peaks with a two-dimensional bottom and the intensity as the third dimension.</p>", "links"=>[], "tags"=>["one-dimensional"], "article_id"=>313907, "categories"=>["Biochemistry", "Infectious Diseases"], "users"=>["Oliver Bannach", "Eva Birkmann", "Elke Reinartz", "Karl-Erich Jaeger", "Jan P. M. Langeveld", "Robert G. Rohwer", "Luisa Gregori", "Linda A. Terry", "Dieter Willbold", "Detlev Riesner"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036620.g001", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_representation_of_the_intensity_distribution_by_an_one_dimensional_scan_through_the_surface_/313907", "title"=>"Schematic representation of the intensity distribution by an one-dimensional scan through the surface.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:37:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/643982"], "description"=>"<p>(<b>A</b>) Image raw data of control sample no. 6 (top) and a scrapie-positive sample no. 7 (bottom). Depicted is one representative out of nine images taken for each sample. Scale bar = 50 µm. (<b>B</b>) Optimization of ‘rolling ball’ radius. After applying rolling ball background subtraction an intensity cutoff (500) was applied. Smaller radii of 1 and 2 pixels (arrows) allow for differentiation of the positive and the control sample (negative). <b>(C)</b> Optimized background removal parameters were applied to the complete panel of samples.</p>", "links"=>[], "tags"=>["Infectious diseases", "Biochemistry"], "article_id"=>314469, "categories"=>["Biochemistry", "Infectious Diseases"], "users"=>["Oliver Bannach", "Eva Birkmann", "Elke Reinartz", "Karl-Erich Jaeger", "Jan P. M. Langeveld", "Robert G. Rohwer", "Luisa Gregori", "Linda A. Terry", "Dieter Willbold", "Detlev Riesner"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036620.g005", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Optimized_image_processing_and_evaluation_/314469", "title"=>"Optimized image processing and evaluation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:40:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/643799"], "description"=>"<p>PrP aggregates from 15 plasma samples were prepared, applied to surface-FIDA using mAb SAF-32 as capture and detection probe and evaluated using a rolling ball radius of 10 px. and intensity cut-off 200. After decoding samples were assigned as scrapie-positive (red) or uninfected controls (green).</p>", "links"=>[], "tags"=>["prp", "particles", "plasma", "scrapie-infected", "blinded"], "article_id"=>314278, "categories"=>["Biochemistry", "Infectious Diseases"], "users"=>["Oliver Bannach", "Eva Birkmann", "Elke Reinartz", "Karl-Erich Jaeger", "Jan P. M. Langeveld", "Robert G. Rohwer", "Luisa Gregori", "Linda A. Terry", "Dieter Willbold", "Detlev Riesner"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036620.g004", "stats"=>{"downloads"=>2, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Detection_of_PrP_particles_in_blood_plasma_of_scrapie_infected_sheep_in_a_blinded_study_/314278", "title"=>"Detection of PrP particles in blood plasma of scrapie-infected sheep in a blinded study.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:39:15"}

PMC Usage Stats | Further Information

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Relative Metric

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