An evaluation of commercial fluorescent bead-based luminex cytokine assays

• Published in: PloS one Vol. 3; no. 7; p. e2535
• Main Authors: Djoba Siawaya, Joel Fleury, Roberts, Teri, Babb, Chantal, Black, Gillian, Golakai, Hawa Jande, Stanley, Kim, Bapela, Nchinya Bennedict, Hoal, Eileen, Parida, Shreemanta, van Helden, Paul, Walzl, Gerhard
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 02.07.2008; Public Library of Science (PLoS)
• Subjects: Analysis; Analytical chemistry; Antigens; Assaying; Biomarkers; Biotechnology; Biotechnology/Protein Chemistry and Proteomics; Blood culture; Blood tests; Cytokines; Cytokines - blood; Dilution; Enzyme-linked immunosorbent assay; Fluorescence; Fluorescent Antibody Technique; Human papillomavirus; Humans; Immunoassay; Immunoassays; Indicators and Reagents - chemistry; Interferon; Manufacturers; Metabolic syndrome; Molecular biology; Multiplexing; Prostate cancer; Reagent Kits, Diagnostic; Sensitivity and Specificity; Viruses; γ-Interferon
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0002535

The recent introduction of fluorescent bead-based technology, allowing the measurement of multiples analytes in a single 25-50 microl sample has revolutionized the study of cytokine responses. However, such multiplex approaches may compromise the ability of these assays to accurately measure actual cytokine levels. This study evaluates the performance of three commercially available multiplex cytokine fluorescent bead-based immunoassays (Bio-Rad's Cytokine 17-plex kit; LINCO Inc's 29-plex kit; and RnD System's Fluorokine-Multi Analyte Profiling (MAP) base kit A and B). The LINCO Inc kit was found to be the most sensitive assay for measuring concentrations of multiple recombinant cytokines in samples that had been spiked with serial dilutions of the standard provided by the manufacturer, followed respectively by the RnD Fluorokine-(MAP) and Bio-Rad 17-plex kits. A positive correlation was found in the levels of IFN-gamma measured in antigen stimulated whole blood culture supernatants by the LINCO Inc 29-plex, RnD Fluorokine-(MAP) and RnD system IFN-gamma Quantikine ELISA kits across a panel of controls and stimulated samples. Researchers should take the limitation of such multiplexed assays into account when planning experiments and the most appropriate use for these tests may currently be as screening tools for the selection of promising markers for analysis by more sensitive techniques.