title: An evaluation of commercial fluorescent bead-based luminex cytokine assays.
abstract: 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.
authors.affiliation: Department of Biomedical Sciences, Division of Molecular Biology and Human Genetics, Stellenbosch University Cape-Town, Stellenbosch, South Africa. joe@sun.ac.za
authors.forename: Joel Fleury
authors.initials: JF
authors.lastname: Djoba Siawaya
authors.forename: Teri
authors.initials: T
authors.lastname: Roberts
authors.forename: Chantal
authors.initials: C
authors.lastname: Babb
authors.forename: Gillian
authors.initials: G
authors.lastname: Black
authors.forename: Hawa Jande
authors.initials: HJ
authors.lastname: Golakai
authors.forename: Kim
authors.initials: K
authors.lastname: Stanley
authors.forename: Nchinya Bennedict
authors.initials: NB
authors.lastname: Bapela
authors.forename: Eileen
authors.initials: E
authors.lastname: Hoal
authors.forename: Shreemanta
authors.initials: S
authors.lastname: Parida
authors.forename: Paul
authors.initials: P
authors.lastname: van Helden
authors.forename: Gerhard
authors.initials: G
authors.lastname: Walzl
mesh_terms: D016207:Cytokines / Q000097:blood*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity
chemical_list: D016207:Cytokines; D007202:Indicators and Reagents; D011933:Reagent Kits, Diagnostic