Blood samples were collected to measure the bone formation markers pro-collagen type 1 N-terminal propeptide (P1NP; Elecsys, Cobas E411, Roche Diagnostics, UK; intraassay %CV <1.7%) and osteocalcin (OCN; Cobas E411, Roche Diagnostics, Germany; intraassay %CV 1.4-3.3%), and the bone resorption marker C-terminal cross-linked telopeptide of type 1 collagen (CTx; Elecsys β-CrossLaps/serum kit, Cobas E411, Roche Diagnostics, UK; intraassay %CV 2.8-8.4%). Factors affecting bone resorption - osteoprotegerin (OPG; ELISA, BioVendor, Czech Republic; intraassay %CV 2.5-4.9%) - and bone formation - sclerostin (SCL; Enzyme Immunoassay, Biomedica Gruppe, Germany; intraassay %CV <7%) - were also measured. Sampling was undertaken as per Figure 1 . All samples were collected in the mornings starting at approximately 08:00 following an overnight fast. Samples were centrifuged, separated and stored at -80°C within 2 hours of collection. Serum samples were assayed in duplicate. P1NP, CTx OPG and SCL were assayed for each sample; OCN only for the baseline and day 8 samples in those receiving 5 days of vibration (insufficient sample for intermediate measures).
Cobas e411
Manufactured by Roche
Sourced in Germany, Switzerland, United States, Japan, United Kingdom, Denmark, France, Poland
The Cobas e411 is a compact and fully automated immunoassay analyzer designed for clinical laboratory settings. It is capable of performing a wide range of immunoassay tests, including those for hormones, therapeutic drug monitoring, and infectious disease markers. The Cobas e411 is known for its reliability, ease of use, and efficient throughput.
Automatically generated - may contain errors
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523 protocols using cobas e411
1
Bone Turnover Markers in Vibration Study
2
Multiplex Biomarker Quantification in Plasma
3
Quantifying SARS-CoV-2 Antibody Levels
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Antibody levels were measured using the Elecsys anti-SARS-CoV-2 S assay (Roche S-RBD tAb). This electrochemiluminescence immunoassay (ECLIA) is used forth in vitro quantification of total antibodies (IgG/IgA/IgM) to SARS-CoV-2 S-RBD proteins inhuman serum and is performed on a Roche Cobas E411 fully automated analyzer (Roche Diagnostics). The assay is performed on the Roche Cobas E411(Roche Diagnostics). This assay is a dual antigen assay format using a recombinant protein representing the RBDS antigen. The three-step procedure favors the detection of high-affinity antibodies to SARS-CoV-2. Samples are incubated with a mixture of biotinylated and ruthenylated RBD antigen to create an immune complex with the dual antigen. Streptavidin-coated microparticles are then added to bind the DAGS complexes to the solid phase. The reagent mixture is transferred to a measuring cell, and the microparticles are trapped magnetically. The application of a voltage induce chemiluminescence, which is measured with a photomultiplier tube. The signal output increases as the antibody titer increases. The detection range is 0.40–250 U/ml (up to 25,000 U/ml at a dilution of 1:100), with values below 0.80 U/ml being considered negative and values above 0.80 U/ml being positive [1 ,16 (link)].
4
Routine Clinical Tests for Patient Evaluation
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Laboratory tests, including routine biochemistry tests and arterial gas analysis, were performed in the hospital laboratory. Blood samples were drawn at admission before the initial therapy, to avoid posttreatment changes in CBC parameters, and collected for differential WBC counts in tubes with EDTA and processed using a Sysmex XN-550 (Sysmex Corporation, Kobe, Japan) automatic blood counting system. Glycated Hb (HbA1c) was measured using a high-performance liquid chromatography kit supplied by Cobas E 411–Roche, Japan. Peptide C was evaluated using automated chemiluminescent assay (Cobas E 411–Roche, Tokyo, Japan). Neutrophil-to-lymphocyte ratios (NLR) were calculated.
5
Bone Biomarker Measurement Protocol
6
Vitamin D and Metabolic Biomarkers Analysis
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Blood samples collected from the subjects during their first visit were analyzed for 25(OH)D and various biochemical parameters. Serum 25(OH)D was determined as described before [20 (link)] with a Roche Elecsys modular analytics (Cobas e411) using an electrochemiluminescence immunoassay (Roche Diagnostics, GmbH, Mannheim, Germany) and commercially available IDS kits (IDS Ltd., Boldon Colliery, Tyne & Wear, UK). Variation for the 25(OH)D ELISA were 5.3% and 4.6%, respectively, with 100% cross-reactivity to 25(OH)D3 and 75% cross-reactivity to 25(OH)D2. It should be noted that the laboratory where samples were analyzed (Biomarkers Research Program, BRP) is a participating entity in the Vitamin D External Quality Assessment Scheme (DEQAS), and Quality Assurance (QA) standards are maintained by ISO 9000 and 17,025. The QA department audits the BRP laboratory at regular intervals. Serum glucose, lipid profile, calcium, and phosphorous were measured using a chemical analyzer (Konelab, Espoo, Finland). Serum-free insulin concentration was determined by electro-chemiluminescence method (Cobas e411; Roche Diagnostics, Mannheim, Germany).
7
Biomarker Panel for Metabolic Health
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Blood (15 mL) and urine were obtained. A total of 5 ml of whole blood were split into two purple top tubes, one frozen for future studies, and the other refrigerated and sent on the same day for HbA1c measurement. The remainder of the blood (10 ml) was centrifuged and the serum was split into several 1 mL Eppendorf tubes. One tube was refrigerated and sent on the same day for glucose measurement. The remaining tubes were kept frozen at − 80 °C for future assays. A fingerstick glucose was obtained at the same time as the laboratory tests.
Laboratory tests were subsequently done using the following methodology: HbA1c by HPLC (Bio-Rad); FPG by Enzymatic method (Cobas 6000, Roche); Serum and Urine Creatinine by the Jaffe rate method (Cobas 6000, Roche); Urine microalbumin by Immunoturbidimetry (Cobas 6000, Roche); TSH, 25(OH) and vitamin D by electrochemiluminescence Immunoassay (ECLIA) (Cobas e 411, Roche), and insulin by immunoassay (Cobas e 411, Roche). Levels of triglycerides, HDL-C, and total cholesterol were measured using enzymatic colorimetric method (Cobas 6000, Roche), and of LDL-C using Coupled Classic precipitation (Cobas 6000, Roche). CRP was measured by Immunoturbidimetric assay (Cobas 6000, Roche).
Laboratory tests were subsequently done using the following methodology: HbA1c by HPLC (Bio-Rad); FPG by Enzymatic method (Cobas 6000, Roche); Serum and Urine Creatinine by the Jaffe rate method (Cobas 6000, Roche); Urine microalbumin by Immunoturbidimetry (Cobas 6000, Roche); TSH, 25(OH) and vitamin D by electrochemiluminescence Immunoassay (ECLIA) (Cobas e 411, Roche), and insulin by immunoassay (Cobas e 411, Roche). Levels of triglycerides, HDL-C, and total cholesterol were measured using enzymatic colorimetric method (Cobas 6000, Roche), and of LDL-C using Coupled Classic precipitation (Cobas 6000, Roche). CRP was measured by Immunoturbidimetric assay (Cobas 6000, Roche).
8
Biochemical and IGFBP-7 Analysis Protocol
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In order to obtain analyzed biochemical parameters and IGFBP 7 concentration, the blood was drawn via a closed system of the Monovette type (Sarstedt Company, Nümbrecht, Germany). The biochemical parameters were determined within 2 h after the material’s collection. Troponin T (TnT) and NT-proBNP analysis was performed using electrochemiluminescence method on the Cobas e411 (ROCHE Diagnostics International Ltd., Rotkreuz, Switzerland). The blood samples (5 mL) used for IFGBP7 determination were left for 2 h at room temperature to allow clot formation. Then, they were centrifuged at 1000× g for 20 min at room temperature, and obtained supernatant serum was frozen and stored at −80 °C. The concentration of IGFBP7 was established with commercially available ELISA kit 7 (insulin growth-factor-binding protein; USCN Life Science Inc., Wuhan, Hubei, China). C-peptide was determined by electrochemiluminescence assay (ECLIA) on Cobas e411 equipment (ROCHE Diagnostics International Ltd., Rotkreuz, Switzerland).
In all participants oral glucose tolerance test (OGTT) and blood pressure and antropometric measurements were performed as described previously [13 (link)].
In all participants oral glucose tolerance test (OGTT) and blood pressure and antropometric measurements were performed as described previously [13 (link)].
9
Assessing Thyroid Function and Immunity in Study
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Following absorption, iodine is metabolized to form thyroid hormones and excess iodine is stored as thyroglobulin [17–19 (link)]. Serum TSH, free T3, and free T4 were measured to assess thyroid hormonal status, whilst thyroglobulin was measured to detect excess circulatory iodine (Cobas e411, Roche, Germany) [17–19 (link)]. Iodine was not measured directly using inductively coupled plasma mass spectrometry for cost considerations. All participants were assessed for anti-SARS-CoV-2 antibodies against the nucleocapsid antigen at baseline and on day 42 (Cobas e411, Roche, Germany).
10
COVID-19 Patient Characteristics and Outcomes
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Patient characteristics, encompassing age, gender, physical examination findings upon admission, GCS score upon admission, co-infection status, and DKA status, were collected. The length of stay in the Intensive Care Unit was determined, along with documentation of critical care interventions, notably the use of mechanical ventilation. Laboratory parameters collected upon admission comprised a CBC, analyzed using a Sysmex XN-550 automated hematology analyzer (Sysmex Corporation, Kobe, Japan), and several biochemistry tests. The latter included C-reactive protein (CRP), fasting glucose level, blood urea nitrogen (BUN), and creatinine, which were assessed using an automatic analyzer (Hitachi 747, Hitachi, Tokyo, Japan). Glycated hemoglobin (HbA1c) was measured via high-performance liquid chromatography (Cobas E 411-Roche, Japan, Tokyo). Additionally, C-peptide was evaluated using an automated chemiluminescent assay (Cobas E 411-Roche, Tokyo, Japan). The SIRI score was calculated based on peripheral blood cell counts using the following formula: (absolute neutrophil count × absolute monocyte count)/absolute lymphocyte count [42 (link)].
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