Blood and urine samples were collected after 8 hours of fasting. All the biochemical analyses were performed in plasma using a COBAS 711 (Roche Diagnostics, Basel, Switzerland), while homocysteine levels were determined using the Immulite 2000 XPi analyser (Siemens Healthcare GmbH, Erlangen, Germany). Haematology values were measured (Sysmex S.L XN 900), and coagulation parameters were determined (ACTLOP analyser).
Immulite 2000 xpi analyser
Manufactured by Siemens
Sourced in Germany
The Immulite 2000 XPi analyser is a fully automated immunoassay analyser designed for clinical laboratory settings. It is capable of performing a wide range of immunoassay tests, including thyroid function, reproductive hormones, and infectious disease markers. The analyser features a compact design, high-throughput capabilities, and user-friendly software interface.
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6 protocols using immulite 2000 xpi analyser
1
Biochemical and Haematology Analyses
2
Analytical Methodologies in Clinical Biochemistry
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All measurements were performed at the department of clinical chemistry and laboratory medicine (AKCL) of Leiden University Medical Center, which is accredited according to CCKL (National Coordination Committee for Quality Assurance for Health Care Laboratories in The Netherlands). Cortisol was measured using an ECLIA assay on a Modular E170 analyser from Roche (Roche Diagnostics, Almere, The Netherlands), ACTH and DHEAS on an Immulite 2000 Xpi analyser (Siemens Healthcare diagnostics, The Hague, The Netherlands) and HbA1c on a Primus Ultra 2 HPLC analyser (Trinity Biotech, Bray, Ireland), using boronate affinity separation. For each participant, all samples from one time series were measured within the same lot number and in the same batch. For this study, the precision and quality of all assayed analytes met or surpassed the level of desirable quality specifications[15 ]. For cortisol Randox controls (Cat. Nr. I/1160EC and 3/1165EC) were used and overall coefficients of variation (CV) for cortisol ranged between 2.4–5.1%, which was well below the desirable CV of 10.5%. For ACTH two levels of controls were used (C2000LACCM1 and C2000LACCM2) and the CV ranged between 3.8–7.7%, which was well below the desirable CV of 10%. In our laboratory the reference range for is ACTH is 3–75 ng/L, for cortisol 0.1–0.6 μmol/L, for HbA1c 20–42 mmol/mol Hb.
3
Growth Hormone Treatment in Nephrotic Syndrome
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Patients with NS treated with GH at a tertiary Children’s Hospital in Liverpool, United Kingdom, were identified. Data was collected retrospectively from the paper medical notes and information was transferred to an electronic database. Research and development approval was obtained (number 6094) and informed consent was not required. Information collected included length of treatment to date and height velocity (HV) for each year on medication. The height measurements were undertaken during the outpatient appointments by trained health care professionals using a standardised stadiometer.
The birth weight standard deviation score (SDS) was corrected to the gestational age. Bone age was analysed using the radius-ulna-short bones method using Tanner-Whitehouse (TW2) method. Both IGF-1 and GH are measured using solid phase, enzyme-labelled, chemiluminescent immunoassays on a Siemens Immulite 2000 XPi analyser [coefficients of variation for low and high levels were 2.82% and 3.80% respectively].
The birth weight standard deviation score (SDS) was corrected to the gestational age. Bone age was analysed using the radius-ulna-short bones method using Tanner-Whitehouse (TW2) method. Both IGF-1 and GH are measured using solid phase, enzyme-labelled, chemiluminescent immunoassays on a Siemens Immulite 2000 XPi analyser [coefficients of variation for low and high levels were 2.82% and 3.80% respectively].
4
Hormonal Biomarker Measurement Protocol
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Serum GH and IGF-1 concentrations were measured by immunoassay using an IMMULITE 2000 Xpi analyser (Siemens). IGF-1 concentrations were normalised for sex and bone age and were expressed as SDS according to the normative data provided by the manufacturer (Siemens Healthcare Diagnostics Inc.). Serum concentrations of TSH, fT4, anti-thyroid peroxidase (anti-TPO) antibodies, and anti-thyroglobulin (anti-Tg) antibodies were measured by immunofluorescence assays using an Architect i1000SR (Abbott Diagnostics).
5
Oral Glucose Tolerance Test Kinetics
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A separate trial was performed to assess plasma glucose and insulin kinetics before (fasting state, time 0) and 30, 50 (corresponding to the start and the end of the haemodynamic measurements, respectively), and 90 min after OGTT. Each participant had an intravenous cannula inserted in a forearm peripheral vein and venous blood (VB) samples collected in pairs: the insulin sample (test tube for serum preparation) immediately after the glucose sample (anticoagulant-coated tube). All test tubes were centrifuged and placed on ice immediately after collection. Serum insulin concentration (cins-VB) was determined by the commercially available solid-phase, enzyme-labelled chemiluminescent immunometric assay, using the “Immulite2000 Insulin” kit and Immulite2000-XPI analyser (both Siemens Healthcare Diagnostics Products Ltd., Camberley, UK). For quantitative determination of plasma glucose (cglc-VB), the enzymatic UV test (hexokinase method) was used (Olympus AU400 system; Mishima Olympus CO., Ltd., Tokyo, Japan; reagent: Beckman Coulter, Inc., Kildare, Ireland).
6
Serum AMH and A4 Quantification
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Blood samples were centrifuged at 1500×g for 10 min within one hour of collection. Hemolytic, lipemic, and icteric serum samples and samples with insu cient volume were excluded from the study. AMH concentrations were assessed by electrochemiluminescence immunoassay method on the UniCel DxI 800 analyser (Beckman Coulter, California, USA). Samples were processed in a single batch according to the manufacturer's instructions. AMH measurement range of the assay was 0.02-24 ng/mL. Internal quality control samples were included in the assay run and intra-assay coe cients of variation (CVs) values were obtained from the measurements. Intra-assay CVs were 1.6% and inter assay CV was 2.6%, for the AMH assay. A4 concentrations were assessed by electrochemiluminescence immunoassay method on Immulite 2000 XPI analyser (Siemens, Eschborn, Germany). Samples were processed in a single batch according to the manufacturer's instructions. A4 analytical sensitivity was 0.3 ng/mL. Internal quality control samples were included in the assay run and intra-assay coe cients of variation (CVs) values were obtained from the measurements. Intra-assay CVs were calculated as 6.4% and inter assay CV was calculated as 7.7.% for the A4 assay.
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