The recent introduction of fluorescent bead-based technology, allowing the measurement of multiples analytes in a single 25–50 µl 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-γ measured in antigen stimulated whole blood culture supernatants by the LINCO Inc 29-plex, RnD Fluorokine-(MAP) and RnD system IFN-γ 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.
This study investigated interleukin-4 (IL-4), IL-4␦2, transforming growth factor beta (TGF-With more than 8 million people progressing to active tuberculosis (TB) every year and a death rate of about 25%, pulmonary TB is one of the most life-threatening human diseases (33). Although the majority of compliant patients are cured after conventional 6-month therapy, about 5% subsequently relapse, mostly within 2 years after completion of treatment. Currently, there is no reliable early predictor of relapse, although failure of sputum smear conversion after the 2-month intensive phase of anti-TB treatment (7, 34) and extent of pulmonary disease on chest radiography (1,7,18,29) have been shown to be risk factors for relapse.
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