Glucose and insulin levels were determined as previously described9 (link) and as surrogate index of the first/early-phase insulin secretion we used the disposition index30 (DI30)9 (link) (details in SI material). High sensitive C-reactive protein (hsCRP) was measured in fasting serum at IPA sampling (1-year follow-up) and yearly during the mean 4-year intervention (active study) period using an IMMULITE® 2000 Systems Analyzer (Siemens Healthcare Diagnostics, Inc. Tarrytown, NY)2 (link). Genotyping of TCF7L2 rs7903146 and rs12255372 was performed as reported10 (link).
Immulite 2000 systems analyzer
Manufactured by Siemens
Sourced in Germany
The IMMULITE® 2000 Systems Analyzers are automated immunoassay analyzers designed for in vitro diagnostic testing. The systems offer random access, continuous loading, and batch processing capabilities for a variety of immunoassay tests.
Automatically generated - may contain errors
Lab products found in correlation
9 protocols using immulite 2000 systems analyzer
1
Genetic Markers, Glucose and Insulin Dynamics
2
Oral Glucose Tolerance Test Protocol
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Glucose and insulin levels were determined as previously described7 (link). In the DPS, during 1993 to 1996 a baseline 2h-OGTT was performed (75 g oral glucose load) with fasting and 2-h samples for glucose and insulin and during follow-up visits starting from the middle of 1996, samples were also taken for 30 min insulin and glucose measurements7 (link). High sensitive C-reactive protein was measured in fasting serum at metabolomics sampling using an IMMULITE® 2000 Systems Analyzer according to the manufacturer instructions (Siemens Healthcare Diagnostics, Inc. Tarrytown, NY).
3
Serum Cortisol Measurement Protocol
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Morning cortisol levels were measured in blood samples taken before food intake between 7:00 and 9:00 am. Blood samples were filled into tubes without additives and centrifuged within 2 h after blood collection. Serum cortisol was quantitatively measured using a chemiluminescent competitive immunoassay on the IMMULITE® 2000 Systems Analyzer (Siemens).
4
Thyroid Autoimmunity Assessment Protocol
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Serum samples were obtained by venipuncture, and blood was collected in 10-mL test tubes. All samples of blood were collected and centrifuged during the routine visits. Serum samples were frozen at -80 °C and stored in a biobank in a specialized clinical laboratory (MCP, Popayan-Colombia). TSH, FT4, anti-TPO, anti-Tg and TRAb were measured using chemiluminescent immunoassay (IMMULITE® 2000 Systems Analyzers; Siemens, Munich, Germany). The coefficients of variation (CVs %) were 4.6%, 6.4%, 4.96%, 8,0% and 8.3% for TSH, FT4, anti-TPO, anti-Tg and TRAb, respectively. The results were classified as follows: TSH: 0.4 - 4.0 mIU/L (normal value); < 0.4 and > 4.0 mIU/L (abnormal value). FT4 levels were only measured in those individuals with a TSH value classified as “abnormal”. The FT4 normal range was 0.89 - 1.76 ng/dL (manufacturer cutoffs). The upper limits of normal for the assays, as denoted by the manufacturers’ reference ranges for diagnosis of thyroid autoimmunity, were used to denote a positive titer for anti-TPO (≥ 35 IU/mL), for anti-Tg (≥ 40 IU/mL) and for TRAb (≥ 0.10 IU/L).
5
Biomarker Measurements in Stored Plasma
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A sample of 30 mL blood was drawn from mainly nonfasting participants at baseline and collected in monovettes containing 10% citrate. Samples were aliquoted and stored in tanks of liquid nitrogen (−196°C) or deep freezers (−80°C) (20 (link)). Plasma NT-proBNP, estradiol, testosterone, and sex hormone–binding globulin (SHBG) concentrations were measured using a solid-phase two-site chemiluminescent immunometric assay (IMMULITE 2000 Systems Analyzers; SIEMENS, Eschborn, Germany) at the Institute of Clinical Chemistry, University of Magdeburg, Magdeburg, Germany. The assay has an effective measuring range of 20–35,000 pg/mL. The median within-run coefficient of variation was 5.4% at a concentration of 35.6 pg/mL and 4.1% at a concentration of 29,725 pg/mL. The overall coefficients of variation throughout the analyses were 6.4% and 4.7% at the same sample concentrations (18 (link)). Plasma adiponectin was measured with a sandwich ELISA (LINCO Research, St. Charles, MO); HDL and total cholesterol, triglycerides, hemoglobin A1c (HbA1c), and hs-CRP were measured using an automatic ADVIA 1650 analyzer (Siemens Medical Solutions) at the University of Tübingen, Tübingen, Germany. All plasma biomarker measurements were corrected for the dilution introduced by citrate volume to improve comparability with concentrations measured in EDTA plasma (22 (link)).
6
Hormonal Profiles in Clinical Study
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Serum total testosterone (TT) and progesterone levels were measured using a solid-phase, competitive chemiluminescent enzyme immunoassay (Immulite 2000 Systems Analyzers, Siemens Medical Solutions Diagnostics, Deerfield, Illinois). The sensitivity for TT was 1.5 ng/mL, and for progesterone was 0.1 ng/mL (0.3 nmol/L). Free testosterone levels were determined by a Coat-A-Count Free Testosterone 125I radioimmunoassay kit purchased from Diagnostic Products Corp. (Los Angeles, California), which had a sensitivity of 0.15 pg/mL. Serum estradiol (E2) levels were measured using a radioimmunoassay kit (Diagnostic Systems Laboratories, Santa Monica, California), and the sensitivity of this assay was 0.22 ng/dL. Serum follicle-stimulating (FSH) levels were measured via enzyme immunoassays (FSH: Abbott Laboratories, Rome, Italy; LH: Dade Behring, Milan, Italy). The sensitivity of FSH assays was 0.2 mIU/mL.
7
Vitamin B12 Deficiency Assessment Protocol
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The blood samples were taken via venipuncture, under fasting conditions; then, they were centrifuged, frozen at −20 °C, and stored. The B12 tests were performed in the serum using a chemiluminescent immunoassay (IMMULITE® 2000 Systems Analyzers; Siemens, Munich, Germany), and the coefficient of variation intra-assay and inter-assay was 7.0% and 6.0%, respectively. The results were classified as follows: <200 pg/mL (deficiency); 200–299 pg/mL (borderline); and ≥300 pg/mL (normal) [8 ,9 (link),10 (link),11 ].
8
Serum Sex Hormone Level Assessment
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Serum total testosterone (TT) and progesterone levels were measured using a solid-phase, competitive chemiluminescent enzyme immunoassay (Immulite 2000 Systems Analyzers, Siemens Medical Solutions Diagnostics, Deerfield, Illinois). The sensitivity for TT was 1.5 ng/mL, and for progesterone was 0.1 ng/mL (0.3 nmol/L). Serum estradiol (E2) levels were measured using a radioimmunoassay kit (Diagnostic Systems Laboratories, Santa Monica, California), and the sensitivity of this assay was 0.22 ng/dL. Serum follicle-stimulating (FSH) and luteinizing hormone (LH) levels were measured via enzyme immunoassays (FSH: Abbott Laboratories, Rome, Italy; LH: Dade Behring, Milan, Italy). The sensitivity for both assays was 0.2mIU/ml. Intra- and interassay coefficients of variations were 4.7% and 8.9%, respectively, for FSH, and 3.1% and 4.0%, respectively, for LH. The sex hormone binding globulin (SHBG) levels were measured using a solid-phase “sandwich” immunoradiometric assay comprising two monoclonal antibodies directed against two different antigen sites, one of which is radiolabeled with iodine125 (Cisbio Bioassays, Codolet, France). This radioimmunoassay had analytical and functional detection limits below 0.5 nmol/L, an intra-assay coefficient of variation (CV) of 5.2%, and inter-assay CVs of 5.3%.
9
Hormonal Analysis Methods Comparison
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Test results were derived from routine testing at the time of test performance. Until September 2014 serum aldosterone was determined by radioimmunoassay Coat-a-Count (Siemens) and later by the automated chemiluminescence immunoassay IDS-iSYS Aldosterone (Immuno Diagnostic Systems). Renin measurement was performed by radioimmunometric assay Renin 3rd Generation (Cisbio) until September 2014 and with the automated chemiluminescence immunoassay IDS-iSYS Direct Renin assay (Immuno Diagnostic Systems). Serum cortisol was analyzed with the chemiluminescent enzyme immunoassay IMMULITE 2000 Systems Analyzers (Siemens).
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