Cardiac c reactive protein latex high sensitive assay

All biomarkers were analysed through commercially available kits. PTX3 was quantified in plasma samples through an enzyme-linked immunosorbent assay (ELISA) kit (Human Pentraxin 3/TSG-14 Quantikine ELISA Kit, R&D Systems, Minnesota, USA). Three classical inflammatory biomarkers were evaluated in serum samples: high-sensitivity (hs) CRP by immunoturbidimetry (Cardiac C-Reactive Protein (Latex) High Sensitive assay, Roche Diagnostics, Basel, Switzerland), IL-6, and TNF-α (Human IL-6 Quantikine HS and Human TNF-alpha Quantikine HS, R&D Systems). Lipid profile, including total cholesterol, triglycerides, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C), was performed using laboratorial routine procedures (Cobas Integra 400 Plus autoanalyser; Roche Diagnostics, Basel, Switzerland); plasma oxidized LDL (oxLDL) and serum adiponectin, and leptin levels were determined through ELISA kits (Oxidized LDL ELISA Kit, Mercodia AB, Uppsala, Sweden; Human Total Adiponectin/Acrp30 Quantikine ELISA Kit and Human Leptin Quantikine ELISA Kit, R&D Systems). Levels of plasma N-terminal pro-B-type natriuretic peptide (NT-proBNP) and serum tissue inhibitor of metalloproteinase-1 (TIMP-1) (Human proBNP and Human TIMP1 ELISA kits, Abcam, Cambridge, UK) were assessed as cardiac and renal fibrosis markers, respectively.

Total leukocyte cell count was assessed in whole-blood using an automatic blood cell counter (Sysmex K1000; Sysmex, Hamburg, Germany).
All inflammatory biomarkers were analyzed through commercially available kits, according to the manufacturer’s instructions. Plasma samples were used to quantify PTX3 (Human Pentraxin 3/TSG-14 Quantikine ELISA Kit, R&D Systems, Minnesota, USA; sensitivity 0.026 ng/mL) and GDF15 (Human GDF15 ELISA Kit, Abcam, Cambridge, UK; sensitivity 2 pg/mL) through enzyme-linked immunosorbent assays (ELISA). In serum we measured IL6 by ELISA (Human IL6 Quantikine HS, R&D Systems; sensitivity 0.09 pg/mL) and hsCRP by immunoturbidimetry [Cardiac C-Reactive Protein (Latex) High Sensitive assay, Roche Diagnostics, Basel, Switzerland; sensitivity 0.15 mg/L].
Genomic DNA was extracted from buffy-coat samples, using genomic DNA extraction kit (GRiSP, Research Solutions, Porto, Portugal), quantified by NanoDrop-1000 (ThermoFisher Scientific, Wilmington, DE, USA) and analyzed by agarose gel electrophoresis. Trademark TaqMan SNP genotyping assays (Human; ThermoFisher Scientific) were performed to assess the allelic frequencies of IL6 (rs1800795) and PTX3 (rs2305619) polymorphisms, using a real-Time PCR system (StepOnePlus, ThermoFisher Scientific).