Plasma insulin was measured using an Abbott AxSYM system (Abbott Laboratories, Abbott Park, IL, USA) by microparticle enzyme immunoassay (Abbott Diagnostics, Wiesbaden, Germany) with an inter-assay coefficient of variation (CV) of <5%. Glucose, triglyceride, total cholesterol, HDL-C, and LDL-C concentrations were measured on a Hitachi 902 autoanalyser (Hitachi High Technologies Corporation, Tokyo, Japan) by enzymatic colorimetric assay (Roche, Mannheim, Germany) with an inter-assay CV of 1.2% for glucose, and <5% for the other parameters.
Microparticle enzyme immunoassay
Manufactured by Abbott
Sourced in United States, Japan, Germany
The Microparticle Enzyme Immunoassay is a laboratory equipment used for the detection and measurement of specific analytes in a sample. It utilizes microparticles coated with antibodies or antigens to capture and quantify the target molecule through an enzyme-based colorimetric reaction.
Automatically generated - may contain errors
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31 protocols using microparticle enzyme immunoassay
1
Metabolic Biomarker Quantification
2
Measurement of Lipid and Inflammatory Biomarkers
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Venous blood from the participants was collected in tubes containing EDTA after a 12-h overnight fast. Lipid variables were assessed with the modular auto analyzer DDPPII Hitachi (Roche, Basel, Switzerland) using specific reagents (Boehringer-Mannheim, Mannheim, Germany). Measurements of total cholesterol (TC) and triglycerides (TG) levels were made by colorimetric enzymatic methods25 (link),26 (link); high-density lipoprotein-cholesterol (HDL-c) levels were measured by colorimetric assay27 (link); and low-density lipoprotein (LDL-C) concentrations were calculated by the Friedewald equation, using the following formula: LDL-C = CT - (HDL-C + TG/5). Glucose measurements were performed using the hexokinase method. hs-C-reactive protein (hs-CRP) concentration was determined by high-sensitivity ELISA (BioCheck, Inc., Foster City, CA, USA). Plasma insulin concentrations were measured by microparticle enzyme immunoassay (Abbott Diagnostics, Matsudo-shi, Japan). Non-esterified fatty acid concentrations were measured by enzymatic colorimetric assay (Roche Diagnostics, Penzberg, Germany). ApoA-1 and ApoB concentrations were determined by immunoturbidimetry.
3
Lipid and Glucose Metabolism Profiling
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Venous blood from the participants was collected in tubes containing EDTA after a 12-hr overnight fast. Lipid variables were assessed with the DDPPII Hitachi modular auto analyzer (Roche, Basel, Switzerland) using specific reagents (Boehringer-Mannheim, Mannheim, Germany). Measurements of total cholesterol (TC) and TG levels were performed by colorimetric enzymatic methods,44 (link), 45 (link) c-HDL was measured by colorimetric assay,46 (link) and low-density lipoprotein (c-LDL) concentration was calculated by the Friedewald equation, using the following formula: c-LDL = CT − (c-HDL + TG/5). Glucose measurements were performed using the hexokinase method. The hs-C-Reactive Protein (hs-CRP) was determined by high-sensitivity ELISA (BioCheck, Foster City, CA, USA). Plasma insulin concentrations were measured by microparticle enzyme immunoassay (Abbott Diagnostics, Matsudo-shi, Japan). Non-esterified fatty acid concentrations were measured by enzymatic colorimetric assay (Roche Diagnostics, Penzberg, Germany). ApoA-1 and ApoB concentrations were determined by immunoturbidimetry.
4
Biomarkers for Atherothrombotic Risk Assessment
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Blood was drawn after a 12-hour fast into EDTA tubes, and plasma samples were stored in freezers at -70°C until analysis. D-dimer levels (nanograms per milliliter) were measured at randomization and at 1 year by microparticle enzyme immunoassay (Abbott Diagnostics) in the MORGAM (MONICA [Multinational Monitoring of Trends and Determinants in Cardiovascular Disease], Risk, Genetics, Archiving, and Monograph) biomarker laboratory. 11 (link) Additional biomarkers in stored plasma samples were chosen to reflect the range of pathobiological processes considered important in the development of atherothrombotic disease: B-type natriuretic peptide (BNP; hemodynamic), sensitive troponin I (myocardial micronecrosis), cystatin C (renal function), high-sensitivity C-reactive protein (hs-CRP; inflammation), lipoprotein-associated phospholipase A 2 activity (inflammation), midregional proadrenomedullin
5
Hepatitis B Serology and Biomarker Assessment
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All serum samples were assessed in each local clinical center laboratory with a standard procedure. Serum HBV surface antigen (HBsAg), HBV surface antibody (HBsAb), HBeAg, HBV e antibody (HBeAb), and HBV core antibody (HBcAb) were measured using microparticle enzyme immunoassay (Abbott Architect, North Chicago, IL). Serum HBV DNA levels were measured using a COBAS TaqMan PCR assay with a lower limit of detection of 20 IU/mL (Roche, Branchburg, NJ). The calculation of FIB-4, APRI, and eGFR were based on the literature.[22 (link),23 (link),24 (link)]
6
Fasting Biomarkers and Inflammation
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Fasting levels of plasma glucose, insulin, and blood lipids were measured using standard laboratory methods (Department of Chemistry, Sahlgrenska University Hospital, Gothenburg, Sweden). LDL cholesterol was calculated using the Friedewald equation [19 (link)]. HbA1c was determined using high-performance liquid chromatography (Mono-S method). In this study, all HbA1c values were converted to International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) standard levels using the formula: HbA1c (IFCC) = (10.45 × HbA1c (Mono-S) -10.62 [20 (link)]. Plasma insulin was measured at the University of Tübingen, Germany, by micro-particle enzyme immunoassay (Abbott Laboratories, Tokyo, Japan).
Total circulating serum adiponectin concentrations were measured by an ultrasensitive ELISA (B-Bridge, Sunnyvale, CA). Plasma IL-6 levels were quantified using an IL-6 specific proliferation bioassay. IL-6 concentrations were calculated using dilutions of recombinant human IL-6 (Genzyme, Cambridge, Massachusetts, USA) as previously described [21 (link)].
Total circulating serum adiponectin concentrations were measured by an ultrasensitive ELISA (B-Bridge, Sunnyvale, CA). Plasma IL-6 levels were quantified using an IL-6 specific proliferation bioassay. IL-6 concentrations were calculated using dilutions of recombinant human IL-6 (Genzyme, Cambridge, Massachusetts, USA) as previously described [21 (link)].
7
Hepatitis B Serological Markers Assay
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Biochemistry was performed using standard laboratory procedures. Hepatitis B surface antigen (HBsAg), antibody to HBsAg (anti-HBs), HBeAg, and antibody to HBeAg (anti-HBe) were measured using microparticle enzyme immunoassay (Abbott Laboratories, North Chicago, IL, USA). Serum HBV DNA levels were measured using a COBAS TaqMan PCR assay (Roche, Branchburg, NJ, USA; lower limit of detection 20 IU/mL).
8
Hepatitis B Serology Profiling
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All serum samples from 210 children were measured for HBsAg and anti-HBc using commercially available ELISA kits (Huakang Biotech, Shenzhen; Kehua Bio-Engineering, Shanghai, China), and were quantitatively tested for antibodies against HBsAg (anti-HBs) with microparticle enzyme immunoassay (Architect system, Abbott, North Chicago), with positive ≥10 mIU/ml. When HBsAg was positive, the sample was further quantitatively tested for HBsAg, HBeAg, and anti-HBc with microparticle enzyme immunoassay (Architect system, Abbott).
9
Fasting Insulin and Glucose Measurements
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Fasting insulin and glucose were measured at the first clinical visit. Fasting insulin (uIU/mL) was measured via Microparticle Enzyme Immunoassay; Abbott Laboratories Diagnostics Division, South Pasadena, CA. Fasting glucose (mg/dL) was measured using Vitros Glucose; Johnson & Johnson; Rochester, NY USA.
10
Cross-Sectional Study of Malaria, Anemia, and Hepcidin in Gambian Children
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All children aged 2–6 years old were recruited from ten rural villages in the West Kiang region of The Gambia during the malaria season (July to August 2001)42 (link). We used cross-sectional data collected at the start of the malaria season. All children had a clinical examination, anthropometric measurements and a 3-d course of mebendazole for possible hookworm infection. A blood sample was collected for complete blood count, observations with a malaria slide, measurements of ferritin (Microparticle Enzyme Immunoassay (Abbott Architect)), hepcidin (Hepcidin-25 (human) EIA Kit (Bachem) and ACT (immunoturbidimetry, Cobas Mira Plus Bio-analyzer, Roche) levels and DNA extraction. Children with a temperature >37.5°C had a malaria blood film, appropriate clinical treatment and a blood sample 2 weeks later after recovery from illness. Genotyping of sickle cell was performed on amplified DNA as detailed elsewhere42 (link). The Gambian Bachem hepcidin values were harmonized by converting to the old DRG hepcidin assay values ((0.266×Bachem values)+1.633) and then to the new High Sensitive DRG hepcidin assay values ((1.989× old DRG values)-3.24) as previously validated43 (link).
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