Performance of Multiplex Commercial Kits to Quantify Cytokine and Chemokine Responses in Culture Supernatants from Plasmodium falciparum Stimulations
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{"title"=>"Performance of Multiplex Commercial Kits to Quantify Cytokine and Chemokine Responses in Culture Supernatants from Plasmodium falciparum Stimulations", "type"=>"journal", "authors"=>[{"first_name"=>"Gemma", "last_name"=>"Moncunill", "scopus_author_id"=>"8951947400"}, {"first_name"=>"John J.", "last_name"=>"Aponte", "scopus_author_id"=>"35594206200"}, {"first_name"=>"Augusto J.", "last_name"=>"Nhabomba", "scopus_author_id"=>"36976170400"}, {"first_name"=>"Carlota", "last_name"=>"Dobaño", "scopus_author_id"=>"6506513895"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"368034725", "sgr"=>"84871760614", "pmid"=>"23300981", "scopus"=>"2-s2.0-84871760614", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0052587", "issn"=>"19326203"}, "id"=>"5b14fde4-d546-3138-8adc-7d568ac39527", "abstract"=>"BACKGROUND: Cytokines and chemokines are relevant biomarkers of pathology and immunity to infectious diseases such as malaria. Several commercially available kits based on quantitative suspension array technologies allow the profiling of multiple cytokines and chemokines in small volumes of sample. However, kits are being continuously improved and information on their performance is lacking. METHODOLOGY/PRINCIPAL FINDINGS: Different cytokine/chemokine kits, two flow cytometry-based (eBioscience(R) FlowCytomix and BD Cytometric Bead Array Human Enhanced Sensitivity) and four Luminex(R)-based (Invitrogen Human Cytokine 25-Plex Panel, Invitrogen Human Cytokine Magnetic 30-Plex Panel, Bio-Rad(R) Bio-Plex Pro Human Cytokine Plex Assay and Millipore MILLIPLEX(R) MAP Plex Kit) were compared. Samples tested were supernatants of peripheral blood mononuclear cells of malaria-exposed children stimulated with Plasmodium falciparum parasite lysates. Number of responses in range that could be detected was determined and reproducibility of duplicates was evaluated by the Bland-Altman test. Luminex(R) kits performed better than flow cytometry kits in number of responses in range and reproducibility. Luminex(R) kits were more reproducible when magnetic beads were used. However, within each methodology overall performance depended on the analyte tested in each kit. Within the Luminex(R) kits, the Invitrogen with polystyrene beads had the poorer performance, whereas Invitrogen with magnetic beads had the higher percentage of cytokines/chemokines with both readings in range (40%), followed by Bio-Rad(R) with magnetic beads (35%). Regarding reproducibility, the Millipore kit had the highest percentage (60%) of cytokines/chemokines with acceptable limits of agreement (<30%), followed by the Invitrogen with magnetic beads (40%) that had tighter limits of agreement. CONCLUSIONS/SIGNIFICANCE: Currently available kits for cytokine and chemokine quantification differ in reproducibility and concentration range of accurate detection. Luminex(R)-based kits with magnetic beads perform the best. Data highlights the importance of testing different kits before each study to choose the most appropriate, depending on the priority of the cytokines assessed.", "link"=>"http://www.mendeley.com/research/performance-multiplex-commercial-kits-quantify-cytokine-chemokine-responses-culture-supernatants-pla", "reader_count"=>68, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>3, "Student > Doctoral Student"=>8, "Researcher"=>14, "Student > Ph. D. Student"=>17, "Student > Postgraduate"=>6, "Student > Master"=>9, "Other"=>2, "Student > Bachelor"=>3, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>3, "Student > Doctoral Student"=>8, "Researcher"=>14, "Student > Ph. D. Student"=>17, "Student > Postgraduate"=>6, "Student > Master"=>9, "Other"=>2, "Student > Bachelor"=>3, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Engineering"=>4, "Environmental Science"=>2, "Nursing and Health Professions"=>1, "Biochemistry, Genetics and Molecular Biology"=>2, "Mathematics"=>1, "Medicine and Dentistry"=>14, "Agricultural and Biological Sciences"=>37, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Immunology and Microbiology"=>3}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>4}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>14}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>37}, "Nursing and Health Professions"=>{"Nursing and Health Professions"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Mathematics"=>{"Mathematics"=>1}, "Unspecified"=>{"Unspecified"=>3}, "Environmental Science"=>{"Environmental Science"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "Netherlands"=>1, "United States"=>2, "China"=>1, "Mexico"=>1, "South Africa"=>1, "Croatia"=>1, "Spain"=>1, "India"=>1}, "group_count"=>4}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/514425"], "description"=>"<p>When differences are <30% the limits of agreement are considered acceptable and are highlighted in bold.</p><p>. (Cytokine/chemokine not tested).</p><p>– (less than 8.1% of both readings in range).</p>*<p>Constant variance p value <0.05.</p>†<p>Constant ratio p value <0.05.</p>‡<p>Ratio is 1 p value <0.05.</p><p>INV-MAG: Invitrogen™ kit with magnetic beads.</p>", "links"=>[], "tags"=>["immunology", "Infectious diseases", "microbiology"], "article_id"=>184920, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Gemma Moncunill", "John J. Aponte", "Augusto J. Nhabomba", "Carlota Dobaño"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052587.t003", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Limits_of_agreement_for_each_cytokine_chemokine_tested_/184920", "title"=>"Limits of agreement for each cytokine/chemokine tested.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-01-02 01:22:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/279432", "https://ndownloader.figshare.com/files/279474", "https://ndownloader.figshare.com/files/279516", "https://ndownloader.figshare.com/files/279561", "https://ndownloader.figshare.com/files/279616", "https://ndownloader.figshare.com/files/279656", "https://ndownloader.figshare.com/files/279704", "https://ndownloader.figshare.com/files/279777", "https://ndownloader.figshare.com/files/279885", "https://ndownloader.figshare.com/files/279964", "https://ndownloader.figshare.com/files/280027", "https://ndownloader.figshare.com/files/280082", "https://ndownloader.figshare.com/files/280153", "https://ndownloader.figshare.com/files/280247", "https://ndownloader.figshare.com/files/280304", "https://ndownloader.figshare.com/files/280374", "https://ndownloader.figshare.com/files/280429", "https://ndownloader.figshare.com/files/280514", "https://ndownloader.figshare.com/files/280568", "https://ndownloader.figshare.com/files/280669", "https://ndownloader.figshare.com/files/280714", "https://ndownloader.figshare.com/files/280801", "https://ndownloader.figshare.com/files/280861", "https://ndownloader.figshare.com/files/280924", "https://ndownloader.figshare.com/files/280965", "https://ndownloader.figshare.com/files/281020", "https://ndownloader.figshare.com/files/281093", "https://ndownloader.figshare.com/files/281163", "https://ndownloader.figshare.com/files/281253", "https://ndownloader.figshare.com/files/281306"], "description"=>"<div><h3>Background</h3><p>Cytokines and chemokines are relevant biomarkers of pathology and immunity to infectious diseases such as malaria. Several commercially available kits based on quantitative suspension array technologies allow the profiling of multiple cytokines and chemokines in small volumes of sample. However, kits are being continuously improved and information on their performance is lacking.</p> <h3>Methodology/Principal Findings</h3><p>Different cytokine/chemokine kits, two flow cytometry-based (eBioscience® FlowCytomix™ and BD™ Cytometric Bead Array Human Enhanced Sensitivity) and four Luminex®-based (Invitrogen™ Human Cytokine 25-Plex Panel, Invitrogen™ Human Cytokine Magnetic 30-Plex Panel, Bio-Rad® Bio-Plex Pro™ Human Cytokine Plex Assay and Millipore™ MILLIPLEX® MAP Plex Kit) were compared. Samples tested were supernatants of peripheral blood mononuclear cells of malaria-exposed children stimulated with <em>Plasmodium falciparum</em> parasite lysates. Number of responses in range that could be detected was determined and reproducibility of duplicates was evaluated by the Bland-Altman test. Luminex® kits performed better than flow cytometry kits in number of responses in range and reproducibility. Luminex® kits were more reproducible when magnetic beads were used. However, within each methodology overall performance depended on the analyte tested in each kit. Within the Luminex® kits, the Invitrogen™ with polystyrene beads had the poorer performance, whereas Invitrogen™ with magnetic beads had the higher percentage of cytokines/chemokines with both readings in range (40%), followed by Bio-Rad® with magnetic beads (35%). Regarding reproducibility, the Millipore™ kit had the highest percentage (60%) of cytokines/chemokines with acceptable limits of agreement (<30%), followed by the Invitrogen™ with magnetic beads (40%) that had tighter limits of agreement.</p> <h3>Conclusions/Significance</h3><p>Currently available kits for cytokine and chemokine quantification differ in reproducibility and concentration range of accurate detection. Luminex®-based kits with magnetic beads perform the best. Data highlights the importance of testing different kits before each study to choose the most appropriate, depending on the priority of the cytokines assessed.</p> </div>", "links"=>[], "tags"=>["multiplex", "kits", "quantify", "cytokine", "chemokine", "responses", "supernatants", "stimulations"], "article_id"=>115093, "categories"=>["Cancer", "Microbiology", "Immunology"], "users"=>["Gemma Moncunill", "John J. Aponte", "Augusto J. Nhabomba", "Carlota Dobaño"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0052587.s001", "https://dx.doi.org/10.1371/journal.pone.0052587.s002", "https://dx.doi.org/10.1371/journal.pone.0052587.s003", "https://dx.doi.org/10.1371/journal.pone.0052587.s004", "https://dx.doi.org/10.1371/journal.pone.0052587.s005", "https://dx.doi.org/10.1371/journal.pone.0052587.s006", "https://dx.doi.org/10.1371/journal.pone.0052587.s007", "https://dx.doi.org/10.1371/journal.pone.0052587.s008", "https://dx.doi.org/10.1371/journal.pone.0052587.s009", "https://dx.doi.org/10.1371/journal.pone.0052587.s010", "https://dx.doi.org/10.1371/journal.pone.0052587.s011", "https://dx.doi.org/10.1371/journal.pone.0052587.s012", "https://dx.doi.org/10.1371/journal.pone.0052587.s013", "https://dx.doi.org/10.1371/journal.pone.0052587.s014", "https://dx.doi.org/10.1371/journal.pone.0052587.s015", "https://dx.doi.org/10.1371/journal.pone.0052587.s016", "https://dx.doi.org/10.1371/journal.pone.0052587.s017", "https://dx.doi.org/10.1371/journal.pone.0052587.s018", "https://dx.doi.org/10.1371/journal.pone.0052587.s019", "https://dx.doi.org/10.1371/journal.pone.0052587.s020", "https://dx.doi.org/10.1371/journal.pone.0052587.s021", "https://dx.doi.org/10.1371/journal.pone.0052587.s022", "https://dx.doi.org/10.1371/journal.pone.0052587.s023", "https://dx.doi.org/10.1371/journal.pone.0052587.s024", "https://dx.doi.org/10.1371/journal.pone.0052587.s025", "https://dx.doi.org/10.1371/journal.pone.0052587.s026", "https://dx.doi.org/10.1371/journal.pone.0052587.s027", "https://dx.doi.org/10.1371/journal.pone.0052587.s028", "https://dx.doi.org/10.1371/journal.pone.0052587.s029", "https://dx.doi.org/10.1371/journal.pone.0052587.s030"], "stats"=>{"downloads"=>100, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Performance_of_Multiplex_Commercial_Kits_to_Quantify_Cytokine_and_Chemokine_Responses_in_Culture_Supernatants_from_Plasmodium_falciparum_Stimulations__/115093", "title"=>"Performance of Multiplex Commercial Kits to Quantify Cytokine and Chemokine Responses in Culture Supernatants from <em>Plasmodium falciparum</em> Stimulations", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-01-02 01:24:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/514476"], "description"=>"a<p>Cost: 1 = highest cost; 4 = lowest cost.</p>", "links"=>[], "tags"=>["immunology", "Infectious diseases", "microbiology"], "article_id"=>184974, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Gemma Moncunill", "John J. Aponte", "Augusto J. Nhabomba", "Carlota Dobaño"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052587.t001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Kits_characteristics_/184974", "title"=>"Kits characteristics.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-01-02 01:22:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/514348"], "description"=>"<p>A) IL-10 and B) IL-2 disagreement plots show the difference between the duplicates against the geometric mean of both values of a sample tested with eBioscience® FlowCytomix™ (FlowCytomix), Millipore™ MILLIPLEX® MAP Plex Kit (Millipore), Bio-Rad® Bio-Plex Pro™ Human Cytokine Plex Assay (Bio-Rad), Invitrogen™ Human Cytokine Magnetic 30-Plex Panel (INV-MAG) and BD™ Cytometric Bead Array Human Enhanced Sensitivity kit (BD CBA). The middle line is the mean difference and the two extreme lines are the limits of agreement (values are truncated to the first decimal) calculated by Bland-Altman test.</p>", "links"=>[], "tags"=>["plots", "il-10", "il-2", "cytokines"], "article_id"=>184842, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Gemma Moncunill", "John J. Aponte", "Augusto J. Nhabomba", "Carlota Dobaño"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052587.g001", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mean_difference_dot_plots_of_IL_10_and_IL_2_cytokines_for_each_kit_tested_/184842", "title"=>"Mean difference dot plots of IL-10 and IL-2 cytokines for each kit tested.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-01-02 01:20:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/514450"], "description"=>"<p>. (not tested).</p><p>Proportions of both readings in range higher than 75% are highlighted in bold.</p><p>INV-MAG: Invitrogen™ kit with magnetic beads.</p>", "links"=>[], "tags"=>["duplicate", "readings"], "article_id"=>184949, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Gemma Moncunill", "John J. Aponte", "Augusto J. Nhabomba", "Carlota Dobaño"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052587.t002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Proportion_of_duplicate_readings_in_range_/184949", "title"=>"Proportion of duplicate readings in range.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-01-02 01:22:29"}

PMC Usage Stats | Further Information

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Relative Metric

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