A Cobas e Immunoassay Analyzer (Roche Diagnostics) was used to determine E2 and β-HCG levels in the follicular fluid according to the manufacturer's protocol. The dilution ratio of follicular fluid ranged from 1:100 to 1:400. The lower and upper limits of E2 detection were 18.4 pmol/l (5 pg/ml) and 11,010 pmol/l (3,000 pg/ml), respectively.
Cobas e immunoassay analyzer
Manufactured by Roche
Sourced in Germany, United States, Switzerland
The Cobas e immunoassay analyzer is a diagnostic instrument manufactured by Roche that is used for automated immunochemistry testing. It is designed to perform various immunoassays, which are analytical techniques used to detect and quantify specific analytes in biological samples such as blood or serum. The core function of the Cobas e analyzer is to automate the process of immunoassay testing, providing accurate and reliable results to support clinical decision-making.
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Lab products found in correlation
Coated tube ria, by DiaSorin (2 mentions)
Autoanalyzer, by Hitachi (2 mentions)
Multi species glp 1 total elisa, by Merck Group (1 mentions)
Statstrip, by Nova Biomedical (1 mentions)
Architect ci8200 analyzer, by Abbott (1 mentions)
Elecsys probnp 2, by Roche (1 mentions)
Cobas vitamin d total assay reagent, by Roche (1 mentions)
Il 6 immunoassay, by Roche (1 mentions)
Eclia, by Roche (1 mentions)
13 protocols using cobas e immunoassay analyzer
1
Follicular Fluid E2 and β-HCG Evaluation
2
Glucose and Hormone Measurements Techniques
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At the Sahlgrenska University Hospital, blood glucose was measured using StatStrip according to the manufacturer’s instructions (Nova Biomedical, Waltham, MA, USA). At Ersta Hospital, blood glucose was measured using the YSI Model 2300 Stat Plus glucose analyzer according to the manufacturer’s instructions (Yellow Springs Instruments, OH, USA). Insulin was measured by an electrochemiluminescence immunoassay “ECLIA” using a Cobas e immunoassay analyzer according to the manufacturer’s instructions (Roche Diagnostics, Dublin, Ireland). The intra-assay variation of the insulin measurement was 1.1–1.4% (CV) and the inter-assay variation 3.5–3.7% (CV). The cross-reactivity with IGF-1 was 0.04%. GLP-1 and GIP were measured using ELISAs according to the manufacturer’s instructions (Merck Millipore, Solna, Sweden; Human total GIP ELISA, product number EZHGIP-54K, and Multi Species GLP-1 Total ELISA, product number EZGLP1T-36K). The intra-assay variation for GLP-1 was 1–2% (CV) and the inter-assay variation < 12% (CV). For GIP, the intra-assay variation was 3–8.8% (CV) and the inter-assay variation 1.8–6.1% (CV).
3
Seasonal Vitamin D and Alkaline Phosphatase
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Blood was collected from the study subjects in the winter season (between January and March) to avoid seasonal variations of vitamin D concentration. Blood samples were collected before 9:00 a.m. on the day of the interview after overnight fasting. All samples were centrifuged within 60 min from blood draw and sera were stored at −70°C until measurement.
The Elecsys Vitamin D(3) (25-OH) immunoassay was used for the quantitative determination of 25-hyroxyvitamin D(3) on cobas e immunoassay analyzer (Roche Diagnostics, Indianapolis, IN, USA). Activity of alkaline phosphatase was measured by colorimetric method (Abbott Laboratories, Abbott Park, Illinois, USA) with para-nitrophenyl phosphate as substrate. The reaction was monitored at 404 nm on an Architect ci8200 analyzer (Architect System, Abbott Laboratories, IL, USA).
Standing height and body weight were measured using standard anthropometric methods (wall-mounted stadiometer, electronic scale; Seca, Germany) and body mass index (BMI) was calculated using the standard formula.
The Elecsys Vitamin D(3) (25-OH) immunoassay was used for the quantitative determination of 25-hyroxyvitamin D(3) on cobas e immunoassay analyzer (Roche Diagnostics, Indianapolis, IN, USA). Activity of alkaline phosphatase was measured by colorimetric method (Abbott Laboratories, Abbott Park, Illinois, USA) with para-nitrophenyl phosphate as substrate. The reaction was monitored at 404 nm on an Architect ci8200 analyzer (Architect System, Abbott Laboratories, IL, USA).
Standing height and body weight were measured using standard anthropometric methods (wall-mounted stadiometer, electronic scale; Seca, Germany) and body mass index (BMI) was calculated using the standard formula.
4
Biomarker Measurement in Pulmonary Hypertension
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During RHC, the fasting blood samples taken from inferior vena cava were collected in EDTA-tubes, centrifuged at 3500× g for 15 min, aliquoted and stored in −80 °C in the Biobank Unit, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Indonesia, until assayed for bio-ADM. An aliquoted sample was thawed, and bio-ADM was measured using a chemiluminescence immunoassay (sphingotest® bio-ADM®, SphingoTec GmbH, Hennigsdorf, Germany) as previously described [6 (link)].
The hemoglobin concentration, hematocrit percentage and N terminal-pro brain natriuretic peptide (NT-proBNP) level (electrochemiluminescence immunoassay (ElecsysProBNP II) and a Cobas e immunoassay analyzer (Roche Diagnostics, Germany)) were measured using blood samples taken during RHC at index of diagnosis in Dr. Sardjito Hospital central laboratory, as previously described [13 (link)].
The hemoglobin concentration, hematocrit percentage and N terminal-pro brain natriuretic peptide (NT-proBNP) level (electrochemiluminescence immunoassay (ElecsysProBNP II) and a Cobas e immunoassay analyzer (Roche Diagnostics, Germany)) were measured using blood samples taken during RHC at index of diagnosis in Dr. Sardjito Hospital central laboratory, as previously described [13 (link)].
5
Thyroid Hormone Measurement by ECLIA
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The serum levels of T3, T4, and TSH were measured by electrochemiluminescence immunoassay “ECLIA” method using Cobas e immunoassay analyzer, using the following Roche Diagnostics GmbH kits: Ref. 11731360 for T3, 06437281 for T4, and 08429324 for TSH. For each kit, the normal expected values correspond to the 2.5th and 97.5th percentile. In our laboratory, the reference ranges for TSH, T4. and T3 were 0.27–4.20 mU/L, 12–22 μg/dL, and 1.3-3.1 ng/dL, respectively.
6
Serum 25(OH)D3 Measurement Protocol
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Blood samples were collected at baseline of this research in RS-I 3rd follow-up, RS-II 1st follow-up and RS-III 1st visit. Serum 25(OH)D3 was assessed using electrochemiluminescence immunoassay (COBAS vitamin D total assay reagent, Roche Diagnostics GmbH, Germany) measured by MODULAR ANALYTICS, Elecsys or Cobas e immunoassay analyzer (Roche Diagnostics GmbH, Germany). The test range was between 7.5 and 175 nmol L−1 with a functional sensitivity of 10 nmol L−1. The intermediate precision of the test was CV < 13.1% and within-run precision was CV < 7.8%. The blood samples were collected at each visit and stored in the freezer at − 80 °C until they were measured for 25(OH)D3 together by the same techniques, instruments and tool kits in the same time period. The concentration of 25(OH)D3 has been adjusted for seasonal variance using cosinor analysis [25 (link)].
7
Elecsys IL-6 Immunoassay for COVID-19
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IL-6 levels were determined in EDTA plasma samples using the commercially available Elecsys IL-6 immunoassay on a Cobas e immunoassay analyzer (Roche Diagnostics, Switzerland), according to manufacturer's instructions. This assay is currently being validated as a predictor of disease progression in COVID-19 and has been granted a US Food and Drug Administration's Emergency Use Authorization (https://diagnostics.roche.com/us/en/products/params/elecsys-il-6.html ). The within-run and between-run precision are 1.9–10.3% and 4.1–11.7%, respectively. Its measuring range is between 1.5–5,000 pg/mL. Normal range is up to 7 pg/ml.
8
Metabolic and Hormonal Biomarkers in PCOS
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Blood glucose, HbA1c, insulin levels, and blood fat, including triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were measured as in a previous population [28 (link)].The electrochemiluminescence immunoassay was performed for measuring serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) by using COBASE immunoassay analyzers (Roche Diagnostics GmbH). An automated analyzer was used for measuring dehydroepiandrostenedione sulfate (DHEA-S) [29 ].
The serum concentration of Fetuin-A was determined by enzyme-linked immunosorbent assay (ELISA) according to the manufacturer’s instructions. Both intra- and inter-assay variations were 10 and 8%, respectively. The measuring range of this kit was 9.38–600 ng/ml. For the determination of sex hormones, venous blood samples in the healthy controls were collected from day 3 to 5 of the menstrual cycle (early follicular period), and in women with PCOS, blood samples were collected after spontaneous bleeding or amenorrhea for more than 3 months [30 (link)].
The serum concentration of Fetuin-A was determined by enzyme-linked immunosorbent assay (ELISA) according to the manufacturer’s instructions. Both intra- and inter-assay variations were 10 and 8%, respectively. The measuring range of this kit was 9.38–600 ng/ml. For the determination of sex hormones, venous blood samples in the healthy controls were collected from day 3 to 5 of the menstrual cycle (early follicular period), and in women with PCOS, blood samples were collected after spontaneous bleeding or amenorrhea for more than 3 months [30 (link)].
9
Automated Elecsys® AMH Assay Evaluation
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About 2 ml of blood was collected in the early follicular phase (cycle day 2–5) for AMH estimation. AMH characteristics in terms of optimal cutoff and area under the receiver operating curve for diagnosing PCOS were estimated on a fully automated Elecsys® and cobas e immunoassay analyzers (Roche Diagnostics GmbH, Germany). Variations of AMH levels in different phenotypes of PCOS were also noted. The test design corresponds to a sandwich immunoassay, based on the streptavidin-biotin technology. The capture antibody is biotinylated; the detection antibody is covalently linked with a Ruthenium complex. Successfully formed antigen-antibody complexes can be detected via electrochemiluminescence within a total assay time of 18 min. The Elecsys® immunoassay detects AMH in the range of 0.01–23 ng/ml (0.07–164 pmol/L) and requires 50 μl of serum or lithium-heparin plasma. The automated Elecsys® AMH assay showed excellent precision, linearity, and functional sensitivity in comparison to the manual AMH assays and showed no interference in the results due to complement binding. Serum AMH levels measured by AMH Gen II are roughly 16% and 20% higher than those obtained with Access AMH and Elecsys AMH, respectively. In addition, serum AMH levels obtained with Elecsys AMH assay are approximately 5% lower than the access AMH assay.[16 (link)]
10
Multiplex Biomarker Quantification Protocol
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Factors were quantified from single samples using a multiplex immunoassay (Proseek Multiplex 96 × 96 CVD III; Olink Bioscience, Uppsala, Sweden), which is a 92-plex immunoassay based on a proximity ligation extension assay. Proximity extension assays use target-specific antibody pairs that are linked to DNA strands that, upon simultaneous binding to the target analyte, create a real-time polymerase chain reaction amplicon in a proximity-dependent manner enabled by the action of a DNA polymerase. The intra-assay coefficient of variation (CV) ranges between 5 and 11% (mean 6%), and the inter-assay CV ranges between 9 and 39% (mean 15%) [8 (link)]. All factors in the assay have been validated and relevant spike-in experiments have been performed to ensure that there is no cross-reactivity between the different biomarkers [51 (link)]. Further information about reproducibility and validation can be found at https://www.olink.com . NT-proBNP levels were analyzed by electrochemiluminescence immunoassay using cobase immunoassay analyzers (Roche Diagnostics, Rotkreuz, Switzerland). Creatinine clearance was calculated according to the Cockcroft–Gault formula.
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