Levels of anti-SARS-CoV2-spike IgG antibodies were measured by an enzyme-linked immunosorbent assay (ELISA) using the Elecsys immunoassay (Roche Diagnostics, Mannheim, Germany) following the manufacturer’s specifications. A positive result was indicated by a value > 0.4 U/mL and the upper measurement limit was set to 2500 U/mL. C-reactive protein (CRP) was measured in serum by an article-enhanced immunological turbidity test, where the aggregates are determined turbidimetrically (Cobas instrument, Roche Diagnostics, Mannheim, Germany). Interleukin-6 (IL-6) was analyzed in plasma samples using the Immunological ECLIA test (ElektroChemiLuminescence ImmunoAssay) on the Cobas instrument (Roche Diagnostics, Mannheim, Germany). Activated Partial Thromboplastin Time (aPTT; reference value: 26–36 s) was measured photometrically (Pathromtin SL, Siemens Healthcare); D-dimers (reference value: <500 µg/L) were analyzed in citrate plasma by a particle-enhanced immunoturbidimetric assay (Innovance D-Dimer Kit, Siemens Healthcare). Anti-β2glycoprotein IgG antibodies and anticardiolipin antibodies were measured by a quantitative ELISA (Euroimmun, Lübeck, Germany). Glucose concentrations were determined in serum using the GLUC3 assay on a Cobas c702 instrument (Roche Diagnostics GmbH, Mannheim, Germany).
Cobas instrument
Manufactured by Roche
Sourced in Switzerland, Germany
The Cobas instrument is a laboratory diagnostic system developed by Roche. It is designed to automate and streamline the analysis of various clinical samples, such as blood, urine, and body fluids. The Cobas instrument utilizes advanced analytical technologies to provide accurate and efficient results, supporting healthcare professionals in making informed decisions.
Automatically generated - may contain errors
Lab products found in correlation
Cobas c702 instrument, by Roche (2 mentions)
Atellica neph 630 system, by Siemens (1 mentions)
Electrochemiluminescent immunoassay, by Roche (1 mentions)
Hd x analyser, by Quanterix (1 mentions)
Au5821 biochemical analyzer, by Beckman Coulter (1 mentions)
Variant 2, by Bio-Rad (1 mentions)
Human galectin 1 quantikine elisa kit, by R&D Systems (1 mentions)
Innovance d dimer kit, by Siemens (1 mentions)
Elecsys immunoassay, by Roche (1 mentions)
Pathromtin sl, by Siemens (1 mentions)
Liaison instrument, by DiaSorin (1 mentions)
Architect instrument, by Abbott (1 mentions)
Tina quant hba1c gen 3 kit, by Roche (1 mentions)
Gluc3 glucose hexokinase kit, by Roche (1 mentions)
Liaison xl analyzer, by DiaSorin (1 mentions)
Variant 2 turbo, by Bio-Rad (1 mentions)
Architect ci16200, by Abbott (1 mentions)
16 protocols using cobas instrument
1
Quantifying COVID-19 Immune Markers
2
Biomarker Measurement in Serum
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C-reactive protein (CRP; reference range 0-0.5 mg/dL) was measured by an article-enhanced immunological turbidity test, where the aggregates are determined turbidimetrically (Cobas instrument, Roche Diagnostics, Mannheim, Germany). Interleukin-6 (IL-6; reference range <15 pg/mL) was measured from serum samples using the Immunological ECLIA test (ElektroChemiLuminescence ImmunoAssay) on Cobas instrument (Roche Diagnostics, Mannheim, Germany). Serum creatinine was analyzed by the Jaffe method. Glucose levels were measured in serum using the GLUC3 assay on a Cobas c702 instrument (Roche Diagnostics GmbH, Mannheim, Germany).
3
Cerebrospinal Fluid Biomarker Measurement
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IgG and albumin concentrations were measured by immunoturbidimetry on a Cobas instrument (Roche Diagnostics). The IgG index and albumin ratio were calculated as previously described.19 (link) Cerebrospinal fluid neopterin was measured using a commercially available immunoassay (Brahms).20 (link) Cerebrospinal fluid β2-microglobulin (β2M) was measured using the N latex β2M kit on the Atellica NEPH 630 System (Siemens Healthcare GmbH). The CSF NfL was measured using a previously described in-house sandwich enzyme-linked immunosorbent assay.21 (link) The NfL concentrations were age adjusted and reported as the equivalent for 65 years of age as previously described.22 (link) The CSF glial fibrillary acidic protein concentration was measured using an in-house enzyme-linked immunosorbent assay as previously described.23 (link) Cytokine concentrations were measured using the novel S-PLEX Proinflammatory Panel I. The S-PLEX format provides signal enhancement that achieves sensitivity in femtograms per milliliters (eMethods in the Supplement ). The CSF and plasma samples were tested undiluted.
4
Plasma NfL and Serum NSE Biomarker Assessment
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All samples were centrifuged, aliquoted, and frozen to −80 °C before storage in the biobank at Region Skane, Sweden (BD-47, SC-1922). The measurements of NfL were performed at the Clinical Neurochemistry Laboratory at the University of Gothenburg in September 2021 by staff blinded to clinical outcomes. Plasma NfL levels were measured using a single-molecule array (Simoa) NfL immunoassay on an HD-X analyser according to instructions from the kit manufacturer (Quanterix, Billerica, MA). Serum neuron-specific enolase (NSE) was analysed by the local laboratory in Region Skane as part of management in clinical practice [12 (link)], using a Cobas instrument with electrochemiluminescent immunoassays (Roche Diagnostics, Rotkreuz, Switzerland).
5
Betatrophin Levels and Metabolic Markers
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Betatrophin levels were measured with the USCNK ELISA kit (Wuhan, China) as directed by the manufacturer, and a full-wavelength microplate reader (SpectraMax Plus 384, MD) was used for absorbance reading. The Cobas instrument (Roche) was used to measure fasting insulin (FINS; 20162404356, Roche Diagnostic GmbH) and 25-(OH)D (20142405153, Roche Diagnostic GmbH) levels by electrochemiluminescence (ECLIA). Then, 75 g OGTT and fasting blood glucose (FBG) level assessment employed the hexokinase method on an AU5821 biochemical analyzer (Beckman Coulter). Hemoglobin A1c (HbA1c) was measured by HPLC (VARIANT-II, Bio-Rad) on a D-10 hemoglobin A1c analyzer (Bio-Rad). To determine whether the inflammatory factor hs-CRP could influence betatrophin levels, immuno-turbidimetric assay was performed on an immage-800 immunoassay system (Beckman Coulter) to measure hs-CRP levels. Homeostasis model assessment (HOMA) was used to calculate insulin resistance index (HOMA-IR) by the following equation: HOMA-IR = FINS × FBG/22.5.
6
Quantifying HIV Biomarkers in Cerebrospinal Fluid
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Cerebrospinal fluid white blood cell (WBC) and blood CD4+ T-lymphocyte counts were measured using routine methods. HIV RNA levels in plasma and CSF were quantified using the Roche Amplicor Monitor version 1.5 or the Roche TaqMan version 1, 2, or 6800 reverse-transcription polymerase chain reaction assays, according to the manufacturer's instructions. The corresponding lower limits of quantification were 50 (Amplicor), 40 (TaqMan v1), and 20 (TaqMan v2 and 6800) HIV RNA copies/mL.
After centrifugation, paired cell-free samples of CSF and plasma were immediately stored at −80°C until later analysis. Archived CSF and plasma samples were subsequently used for all study analyses.
CSF and serum neopterin concentrations were measured using a commercially available immunoassay (BRAHMS, Henningsdorf, Germany) with an upper normal reference value of 5.8 nmol/L in CSF and 9.1 nmol/L in serum [6 (link), 13 ]. Immunoglobulin G (IgG) concentrations were measured by immunoturbidimetry on a Cobas instrument (Roche Diagnostics, Penzberg, Germany). IgG index was calculated as previously described [14 (link)]. CSF β2-microglobulin (β2M) was measured using the N Latex β2M kit on the Atellica NEPH 630 System (Siemens Healthcare GmbH. Erlangen, Germany).
After centrifugation, paired cell-free samples of CSF and plasma were immediately stored at −80°C until later analysis. Archived CSF and plasma samples were subsequently used for all study analyses.
CSF and serum neopterin concentrations were measured using a commercially available immunoassay (BRAHMS, Henningsdorf, Germany) with an upper normal reference value of 5.8 nmol/L in CSF and 9.1 nmol/L in serum [6 (link), 13 ]. Immunoglobulin G (IgG) concentrations were measured by immunoturbidimetry on a Cobas instrument (Roche Diagnostics, Penzberg, Germany). IgG index was calculated as previously described [14 (link)]. CSF β2-microglobulin (β2M) was measured using the N Latex β2M kit on the Atellica NEPH 630 System (Siemens Healthcare GmbH. Erlangen, Germany).
7
Quantifying Galectin-1 and Cytokines
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Galectin-1 was measured using the Human Galectin-1 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturers' instructions. Intra-assay and inter-assay coefficient of variation were 7.1% and 9.5%, respectively. Cytokines were measured in serum using the ultrasensitive multiplex electrochemiluminescence immunoassay (ELISA) (Meso Scale Diagnostics (MSD), Rockville, MD, USA). The human V-PLEX Pro-Inflammatory Panel 1 Human Kit (#K15049) was used to quantify interferon gamma (IFN-γ), interleukin (IL)-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13 and tumor necrosis factor-alpha (TNF-α) according to the manufacturer's instructions.
Routine analyses of glucose, insulin, and cholesterol were performed at the Department of Clinical Chemistry, Sahlgrenska University Hospital, on a Cobas instrument (Roche Molecular Diagnostics). The homeostasis model assessment for insulin resistance (HOMA-IR) was calculated using the formula [glucose]*[insulin]/22.5 [7 (link)].
Routine analyses of glucose, insulin, and cholesterol were performed at the Department of Clinical Chemistry, Sahlgrenska University Hospital, on a Cobas instrument (Roche Molecular Diagnostics). The homeostasis model assessment for insulin resistance (HOMA-IR) was calculated using the formula [glucose]*[insulin]/22.5 [7 (link)].
8
Serum Biomarkers in Type 2 Diabetes
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After obtaining proper informed consent for exploratory analyses, baseline serum samples were collected from 93 patients with type 2 diabetes enrolled in a clinical trial. These samples were drawn under fasting morning conditions using serum separator tubes and stored at -80 °C prior to subsequent analyses. Serum samples were also obtained from 99 normal subjects for the Eli Lilly Research Blood Donor Program after the anonymized subjects gave their informed consent for sample collection. These samples were also drawn under fasting morning conditions using serum separator tubes and stored at -80 °C prior to subsequent analyses. All lipid markers and ApoB were measured using a Roche Cobas instrument. The ratio of ApoB to low density lipoprotein cholesterol (LDL-C) was used as a surrogate marker for small dense LDL-C. Non-HDL-C was calculated by subtracting HDL-C from total cholesterol (TC). Levels of TG and HDL-C were used to calculate the TG/HDL ratio.
9
Fasting Blood and Urine Analysis for Hormone Profiles
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Overnight fasting blood samples were collected at baseline and 1.5, 3, and 6 months of intervention. Automated biochemical profiles were measured at the Biomedical Diagnostic Center of the Hospital Clinic (Table 2 ).
Blood from each visit was drawn into ethylenediaminetetraacetic acid (EDTA) collection tubes, and plasma was separated after centrifugation at 1500 g (RCF) for 15 min at 7 °C. 24-h urine samples were also collected at all visits. Plasma and 24-h urine samples were stored in aliquots at −80 °C until the day of analysis.
Stored plasma aliquots collected at the different time points were used to analyse sex hormones. FSH, LH, progesterone, E2, and sex hormone-binding globulin (SHBG) were measured by a chemiluminescent immunoassay using an Atellica instrument (Siemens), while total testosterone (T-total) was measured by a direct chemiluminescent immunoassay with a Cobas instrument (Roche). The free testosterone index (FTI) was defined as the ratio between testosterone levels and SHBG levels, multiplied by a constant. To study the bioavailable E2, the free oestradiol index (FEI) was calculated as the molar ratio of plasma E2 to the plasma SHBG level and multiplying by 100 [30 (link)]. The lower detection limits of plasma E2 and progesterone were 12 pg/mL and 0.21 ng/mL, respectively; levels below these limits were defined as 11 pg/mL of E2 and 0.20 ng/mL of progesterone.
Blood from each visit was drawn into ethylenediaminetetraacetic acid (EDTA) collection tubes, and plasma was separated after centrifugation at 1500 g (RCF) for 15 min at 7 °C. 24-h urine samples were also collected at all visits. Plasma and 24-h urine samples were stored in aliquots at −80 °C until the day of analysis.
Stored plasma aliquots collected at the different time points were used to analyse sex hormones. FSH, LH, progesterone, E2, and sex hormone-binding globulin (SHBG) were measured by a chemiluminescent immunoassay using an Atellica instrument (Siemens), while total testosterone (T-total) was measured by a direct chemiluminescent immunoassay with a Cobas instrument (Roche). The free testosterone index (FTI) was defined as the ratio between testosterone levels and SHBG levels, multiplied by a constant. To study the bioavailable E2, the free oestradiol index (FEI) was calculated as the molar ratio of plasma E2 to the plasma SHBG level and multiplying by 100 [30 (link)]. The lower detection limits of plasma E2 and progesterone were 12 pg/mL and 0.21 ng/mL, respectively; levels below these limits were defined as 11 pg/mL of E2 and 0.20 ng/mL of progesterone.
10
Serum Biomarkers for Bone Health
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Serum samples were frozen at −80 °C immediately after collection and stored for up to 1 year before analysis. After thawing, the samples were all analyzed on the same day.
Serum 25(OH)D (both D2 and D3) and serum osteocalcin concentrations were analyzed with chemiluminescence immunoassay (CLIA) on a LIAISON instrument (DiaSorin Inc, Stillwater, MN, USA).
The total coefficient of variance (CV) for serum 25(OH)D was 5–6 %, with the highest variance in the lowest test range, and the functional sensitivity was 12.5 nmol/L at a CV of 8 %.
The total CV for serum osteocalcin was 4–6.5 %, with the highest variance in the lowest test range, and the functional sensitivity was 3 µg/L at a CV of 17 %.
Serum concentrations of intact parathyroid hormone (iPTH) were analyzed with CLIA on an Abbott ARCHITECT instrument (Abbott Diagnostics Division, Abbott Park, IL, USA). The total CV for iPTH ranged from 2.8 to 3.2 %. The functional sensitivity was below 5 ng/L at a CV of 20 %. The reference interval for iPTH, provided by the manufacturer, was 15–68 ng/L (percentile 2.5–97.5).
Serum calcium, albumin, and phosphate were analyzed on a Cobas instrument (Roche Molecular Diagnostics, Pleasanton, CA, USA).
Serum 25(OH)D (both D2 and D3) and serum osteocalcin concentrations were analyzed with chemiluminescence immunoassay (CLIA) on a LIAISON instrument (DiaSorin Inc, Stillwater, MN, USA).
The total coefficient of variance (CV) for serum 25(OH)D was 5–6 %, with the highest variance in the lowest test range, and the functional sensitivity was 12.5 nmol/L at a CV of 8 %.
The total CV for serum osteocalcin was 4–6.5 %, with the highest variance in the lowest test range, and the functional sensitivity was 3 µg/L at a CV of 17 %.
Serum concentrations of intact parathyroid hormone (iPTH) were analyzed with CLIA on an Abbott ARCHITECT instrument (Abbott Diagnostics Division, Abbott Park, IL, USA). The total CV for iPTH ranged from 2.8 to 3.2 %. The functional sensitivity was below 5 ng/L at a CV of 20 %. The reference interval for iPTH, provided by the manufacturer, was 15–68 ng/L (percentile 2.5–97.5).
Serum calcium, albumin, and phosphate were analyzed on a Cobas instrument (Roche Molecular Diagnostics, Pleasanton, CA, USA).
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