Blood samples were collected in the early morning after overnight fasting. In all cases, 30 ml of blood was extracted by venipuncture from the antecubital vein by a qualified nurse. The stability of samples during transport and storage in the HELENA study has been reported previously (38 ). Nine key biomarkers involved in low-grade, systemic inflammation were selected for this study (CRP, TNF-α, IL-6, 1,2,4,10, IFN-γ, sICAM and sVCAM. CRP was measured in serum by immunoturbidimetry (AU2700 biochemistry analyzer; Olympus, Watford, UK). Serum cytokines IL-6, 1, 2, 4, 10 and TNF-α were determined using the High Sensitivity Human Cytokine MILLIPLEXTM MAP kit (Millipore Corp., Billerica, MA, USA) and collected by flow cytometry (Luminex-100 v.2.3, Luminex Corporation, Austin, TX, USA). The intra- and inter-assay precision CVs were: 3.5% and 4.5%, respectively, for IL-6; and 3.5% and 3.8%, respectively, for TNF-α.
Au2700 biochemistry analyzer
Manufactured by Olympus
Sourced in United Kingdom
The AU2700 is a biochemistry analyzer produced by Olympus. It is designed to perform a variety of clinical chemistry tests on biological samples, such as blood and urine. The AU2700 utilizes spectrophotometric technology to analyze the components of a sample and provide quantitative results.
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Lab products found in correlation
5 protocols using au2700 biochemistry analyzer
1
Biomarkers of Systemic Inflammation Assessment
2
Biomarker assessment in coronary heart disease
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Blood samples were obtained for research purposes, with informed consent from the subjects, after an overnight fast of at least 10 h. The tested analytes were obtained by spectrophotometry using an Olympus AU 2700 (Olympus Diagnostic, Hamburg, Germany). The hs-CRP levels were determined by the immunoturbidimetric method applied via the Olympus AU 2700 biochemistry analyzer. The regents and calibrators were used according to the manufacturers’ recommendations with analytic measuring ranges of 0.05–20 mg/dL for the hs-CRP test. Finally, the NT-pro-BNP levels were measured by immunoassay using the Siemens Stratus CS Acute Care Diagnostic System (Siemens Healthcare Diagnostics, Inc., Newark, DE, USA). Biochemistry analyzers and the same reference values were used for all the CHD and control patients.
3
Measuring Exhaled Nitric Oxide and Insulin Sensitivity
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To obtain FeNO samples, the participants were asked to inhale through activated carbon to remove ambient nitric oxide and to exhale into individual aluminum bags at a flow rate of 150 L/h at a positive pressure of . The nitric oxide concentrations in the samples were analyzed using a calibrated chemiluminescence nitrogen oxide analyzer (Model 42i; Thermo Scientific). Serum levels of FBG and fasting plasma insulin (FINS) were measured with an Olympus AU2700 biochemistry analyzer at Peking University Hospital. HOMA-IR was calculated using the levels of FBG and FINS ( ) to estimate the basal insulin sensitivity (Levy et al. 1998 (link)). Weight was measured for each participant using a weighing scale (HBF-358-BW; Omron Healthcare, Inc.), and height was measured using a meter stick. Body mass index (BMI) was calculated as the weight divided by the square of the height.
4
Blood Collection and Biochemical Analysis
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At 8:00 A.M. after a 12-hour overnight fast, 10 ml of blood was extracted by venipuncture in the antecubital fossa of the right arm with a disposable vacuum blood collection tube. In the four hours after the extraction, all samples were centrifuged at 1300g for 15 minutes (Z400 K, Hermle, Wehingen, Germany). The red blood cells were thus separated and the serum was finally frozen at -80ºC for its subsequent analysis. Immediately after collection and before centrifugation, however, we measured glucose concentration using an enzymatic colorimetric method (glucose oxidase-phenolaminophenazone [GOD-PAP] method, Human Diagnostics, Germany) as well as the concentrations of HDL-C, total Newwark, USA; R&D Systems, Minneapolis, USA). hs-CRP was determined by solid-phase chemiluminiscent immunometric assay. Ceruloplasmin was measured by immunoturbidimetry (AU2700 biochemistry analyzer; Olympus). Serum insulin was determined by radioimmunoanalysis (Insulin Kit, DPC, Los Angeles, EEUU). Insulin resistance was quantified with HOMA (Homeostasis Model Assessment) [20] by applying the following formula: fasting glucose (mmol/L) x fasting insulin (mU/L)/22.5.
5
Standardized Fasting Blood Sampling Protocol
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In a randomly selected one-third subset of the total HELENA study population, blood samples were collected in overnight fasting state. We used a standardized methodology for blood collection, transport and analysis by a certified laboratory [31] (link). The quality control was within recommended levels and transport had no influence. Detection limits (sensitivity) for serum CRP by immunoturbidimetry (AU2700 biochemistry analyzer, Olympus, Watford, UK) were 0.007 mg/L and 0.05 pg/mL with intra-assay CV of 1.9%. Nobody took nonsteroidal anti-inflammatory drugs or had an active inflammatory disease. None of the girls had Polycystic Ovarian Syndrome or was in the menstrual period. Six individuals (only male) presented CRP levels higher than 10 mg/L. Only 8.4% smoked regularly.
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