Quantikine human

Serum samples were obtained after 8–10 h overnight fasting, and frozen immediately at −80°C. Routine biochemical parameters were measured using standard methods. Serum high sensitive C-reactive protein (hs-CRP) was measured by an immunoturbidimetric fully automated assay (Roche Diagnostics GmbH, Mannheim, Germany) with functional sensitivity of 0.3 mg/L (intra- and inter-assay precision, 1.6 and 8.4, resp.). Serum levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and IL-10 were determined by a high-sensitive ELISA method (Quantikine Human, R&D Systems, Minneapolis, USA) in a DSXTM 4 Plate ELISA Processor (Vitro SA, Spain). Minimum detectable concentrations were 0.10 pg/mL, 0.70 pg/mL, and 0.50 pg/mL, respectively. Intra- and inter-assay coefficients of variation (CV) were <10.8% in all cases.