Performance of multiplex commercial kits to quantify cytokine and chemokine responses in culture supernatants from Plasmodium falciparum stimulations

• Published in: PloS one 2013-01, Vol.8 (1), p.e52587-e52587
• Main Authors: Moncunill, Gemma, Aponte, John J, Nhabomba, Augusto J, Dobaño, Carlota
• Format: Article
• Language: English
• Subjects: Beads; Biology; Biomarkers; Biomarkers - metabolism; Cell culture; Chemokines; Chemokines - metabolism; Child; Children; Cytokines; Cytokines - metabolism; Cytometry; Erythrocytes - parasitology; Flow Cytometry; Hospitals; Humans; Immunity; Immunoassay; Infectious diseases; Interleukin-10 - blood; Interleukin-2 - blood; Leukocytes (mononuclear); Leukocytes, Mononuclear - cytology; Lysates; Magnetics; Malaria; Malaria - diagnosis; Medical research; Medicine; Methods; Multiplexing; Pathology; Peripheral blood mononuclear cells; Plasmodium falciparum; Plasmodium falciparum - metabolism; Polystyrene; Polystyrene resins; Reagent Kits, Diagnostic - standards; Reproducibility; Reproducibility of Results; Sensitivity enhancement; Studies; Time Factors; Vector-borne diseases
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0052587

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. 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 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® 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 (