Euroline systemic sclerosis nucleoli profile igg

Antinuclear antibodies (ANA) were evaluated by indirect immunofluorescence (IIF) assay using HEp-cell line 2. Anticentromere antibodies (ACA) were described by IIF, and anti-topoisomerase I antibodies were determined by enzyme-linked immunosorbent assay. A commercial line blot assay was performed to detect anticentromere proteins A and B, anti-RNA pol III and anti-PM/Scl antibodies (EUROLINE Systemic Sclerosis (Nucleoli) Profile (IgG), Euroimmun, Germany) according to the manufacturer’s instructions.

All patients were tested for LA, aCL (IgG isotype) and anti-β2GpI (IgG isotype). LA was detected in plasma by a dilute Russell's viper venom time (Siemens), and partial thromboplastin time test (HemosIL Silica Clotting Time Werfen) as screening and confirmation tests with calculating a normalized ratio. aCL and anti-β2GpI were measured using commercial ELISA assays (Orgentec, Trappes, France), positive titer was defined as ≥10 UGPL/mL (aCL) or ≥10 UA /mL (anti-β2GpI). Identification of antinuclear antibody specificities using both specific immunofluorescence patterns on HEp-2 cells and the Luminex approach (Bio-Plex 2200; Bio-Rad) for anti–topo I, ACA, anti–U1 RNP, anti-SSA/Ro, and anti-SSB/La antibodies was performed as part of routine clinical care. Anti–RNA pol III antibodies were identified by immunodot (Euroline Systemic Sclerosis [Nucleoli] Profile [IgG]; Euroimmun). Other laboratories tests performed at the inclusion were: creatinine, CRP, platelet count, uric acid, serum protein electrophoresis, immunoglobulin G, M, and A plasma levels, LDL-cholesterol, triglycerides and glycated hemoglobin. Diabetes mellitus was defined as a glycated hemoglobin ≥6.5% and/or anti-diabetic medication intake. Dyslipidemia was defined as a LDL-cholesterol ≥1.6 g/L and/or triglycerides ≥1.5 g/L and/or lipid-lowering medication intake.

The presence of antinuclear antibodies (ANA) was evaluated by indirect immunofluorescence on HEp-2000 cells-coated slides (Eurobio Ingen, Les Ulis, France) with a positivity threshold set at a titer of 1:100. ANA specificities were then determined by performing both a multiplex fluorescent microsphere immunodetection (FIDIS Connective Profile MX117™, Theradiag^®, Croissy-Beaubourg, France) and an immunodot assay (Euroline Systemic Sclerosis [Nucleoli] Profile [IgG]; Euroimmun, Luebeck, Germany), allowing separate detection of anti-DNA topoisomerase I, anti-centromere AAb, anti–RNA polymerase III, anti-fibrillarin, anti-NOR90, anti Th/To, anti-PM-Scl, anti-Ku, anti-PDGFR, and anti-Ro-52 antibodies.