Enzyme immunoassay test kit

A full joint examination was performed by the same paediatric rheumatologist during the outpatient visit. Blood samples were obtained for nine different biochemical markers: glucose, creatinine, total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, triglycerides, ADMA, adiponectin and MPO. The first six markers are regularly used in clinical practice and were analysed in the local clinical laboratory using certified standard assays. For the last three markers, serum and plasma were stored at −80 °C until analysis. Serum ADMA concentration was determined by using a validated enzyme-linked immunosorbent assay (ELISA) kit (DLD Gesellschaft für Diagnostika und medizinische Geräte mbH, Hamburg, Germany). Serum adiponectin concentration was determined using the Human Adiponectin/Acrp30 Quantikine ELISA kit (R&D Systems Europe, Abingdon, UK). The adiponectin intraassay coefficient of variation (CV) is 4.6 %, and the interassay CV is 7.9 %. MPO was determined in serum of patients with JIA using an enzyme immunoassay test kit (catalogue number BC-1129; BioCheck, Foster City, CA, USA) and in plasma of controls using an ELISA kit (BIOXYTECH enzyme immunoassay for MPO, catalogue number 21013; OXIS Health Products, Portland, OR, USA), according to the manufacturer’s instructions. MPO intraassay CV is 5.7 % and inter-assay CV 8.5 %.

5 cc venous blood was collected from every case using standard venipuncture aseptic technique. 1 cc taken on EDTA tube for complete blood picture assessment and the rest was centrifuged and sera were collected and stored at -20^℃ for assessment of serum adiponectin, serum resistin, serum MCP-1 & total serum IgE using Enzyme linked immuno-sorbent assay (ELISA) technique.

• Serum IgE (immunoglobulin E): enzyme immunoassay test kit provided by Bio Check, Inc.; Foster city, CA 94404, USA; for the quantitative determination of IgE concentration in human serum)

• Serum resistin: Enzyme-linked immunosorbent assay kit, for quantitative detection of human resistin, provided by Assaypro LLC 30 Triad South Drive St. Charles, MO 63304, USA)

• Serum adiponectin: ELISA kit, for quantitative sandwich enzyme immunoassay technique that measures adiponectin in less than 3 hours, provided by Assaypro LLC 30 Triad South Drive St. Charles, MO 63304, USA).

• Serum MCP-1: ELISA kit, for quantitative determination of human MCP-1 in human serum provided by glory science co., ltd. 2400 veterans Blvd. Suite 16 – 101, Del Rio, TX 78840, USA).