Performance of Luminex and MSD high-sensitivity multiplex cytokine assays across multiple lots and laboratories (65.6)

Abstract

Abstract Circulating cytokine levels can provide valuable information about immune status, but often require high-sensitivity assays. Multiplex assays hold great promise, especially when limited sample volumes are available. Four high-sensitivity cytokine multiplex assays (BioRad, BioSource/Invitrogen, Linco/Millipore, Meso Scale Discovery [MSD]) were evaluated for ability to detect circulating levels of cytokines, and for lot and laboratory variability. Assays were performed in six different laboratories utilizing archived serum from HIV-uninfected and -infected subjects from the Multicenter AIDS Cohort Study (MACS) and the Women’s Interagency HIV Study (WIHS), and commercial plasma samples spanning initial HIV viremia. In a majority of serum samples IL6, IL8, IL10, and TNFα were detectable with at least three kits, while IL1β was clearly detected with only one kit. No single multiplex panel detected all cytokines, and there were highly significant differences (p<0.001) between laboratories and/or lots with all kits. The kits generally detected similar cytokine patterns over the onset of HIV viremia. This multi-site comparison suggests that current multiplex assays vary in their ability to measure serum and/or plasma levels of cytokines, and may not be sufficiently reproducible for repeated determinations over a long-term study or in multiple laboratories, but may be useful for longitudinal studies in which relative, rather than absolute, changes in cytokines are important.