Blood was collected from the 51 subjects to measure biochemical values at week 0 (before the experiment), and the 4th, 12th, 24th, 48th weeks (in the experiment) and in the 4th and 12th weeks after the experiment. Heparinized blood was collected to evaluate liver and kidney function and genotype, and to quantify HBV-DNA concentration, HBsAg, and HBeAg. Serum HBV-DNA levels (copies/mL) were measured by polymerase chain reaction (PCR) using Cobas TaqMan HBV Test, v2.0 (Roche Molecular Systems, Pleasanton, CA, USA), according to the manufacturer’s instructions. Quantification of HBsAg was performed by an automated chemiluminescent micro-particle immunoassay (CMIA) using the Roche Cobas e602 analyzer with Elecsys HBsAg II Quant reagent kits (Roche Diagnostics, Santa Clara, CA, USA), according to the manufacturer’s instructions. Quantification of HBeAg was conducted by an automated chemiluminescent micro-particle immunoassay (CMIA) using the Roche Cobas e602 analyzer with Elecsys HBeAg Quant reagent kits (Roche Diagnostics, CA, USA), according to the manufacturer’s instructions. The remaining blood was segregated into serum and cells. Serum was stored as aliquots in liquid nitrogen until analysis and cells were analyzed to determine the T cell subtypes.
Cobas e602 analyzer
Manufactured by Roche
Sourced in United States, Switzerland, Germany, Singapore
The Cobas e602 analyzer is a fully automated immunoassay analyzer designed for the in vitro determination of various analytes in human samples. The core function of this instrument is to perform immunoassay tests, which are used to detect and measure the presence of specific substances in the body, such as hormones, proteins, and antibodies.
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35 protocols using cobas e602 analyzer
1
Longitudinal Analysis of HBV Markers
2
Standardized Biomarker Measurement in Serum
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All expressed biomarkers were measured in the serum of patients’ blood. All samples were obtained by venipuncture into serum monovettes® and centrifuged at 2000 g for 10 min at 20 °C. The aliquoted samples were cooled down with liquid nitrogen before being stored at − 80 °C until analysis. The complete processing was conducted within two hours after blood extraction. After thawing, the samples were mixed gently by inverting and centrifuged with 2500g for 10 min at 20 °C, respectively, 3000g for 30 min for hsTnI at 4 °C.
HsTnT was measured with the Troponin T hs STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The limit of blank (LoB) for this assay was 3 ng/L and the limit of detection (LoD) was 5 ng/L as described in the instructions for use [15 ]. HsTnI was measured with the STAT High sensitivity Troponin-I assay on an Architect i1000 analyzer (Abbott, Wiesbaden, Germany). The LoB was 0.7–1.3 ng/L and the LoD was 1.1–1.9 ng/L for this assay as described in the instructions for use [16 ]. NT-proBNP was measured with the proBNP II STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The LoD for this assay was 5 ng/L [17 ]. Creatinine was measured with the Creatinine Jaffe Gen.2 assay on a cobas c 702 analyzer (Roche Diagnostics, Mannheim, Germany).
HsTnT was measured with the Troponin T hs STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The limit of blank (LoB) for this assay was 3 ng/L and the limit of detection (LoD) was 5 ng/L as described in the instructions for use [15 ]. HsTnI was measured with the STAT High sensitivity Troponin-I assay on an Architect i1000 analyzer (Abbott, Wiesbaden, Germany). The LoB was 0.7–1.3 ng/L and the LoD was 1.1–1.9 ng/L for this assay as described in the instructions for use [16 ]. NT-proBNP was measured with the proBNP II STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The LoD for this assay was 5 ng/L [17 ]. Creatinine was measured with the Creatinine Jaffe Gen.2 assay on a cobas c 702 analyzer (Roche Diagnostics, Mannheim, Germany).
3
Biomarker Quantification in Serum Samples
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All expressed biomarkers were measured in the serum of patients' blood. All samples were obtained by venipuncture into serum monovettes Õ and centrifuged at 2500 g for 10 min at 20 C. The aliquoted samples were cooled down with liquid nitrogen before being stored at À80 C until analysis. The complete processing was conducted within 2 h after blood extraction. After thawing, the samples were mixed gently by inverting and centrifuged with 2500 g for 10 min at 20 C, respectively, 3000 g for 30 min for hsTnI at 4 C.
HsTnT was measured with the Troponin T hs STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The limit of blank (LoB) for this assay was 3 ng/L and the limit of detection (LoD) was 5 ng/L as described in the instructions for use. 29 HsTnI was measured with the STAT High sensitivity Troponin-I assay on an Architect i1000 analyzer (Abbott, Wiesbaden, Germany). The limit of blank (LoB) was 0.7-1.3 ng/L for this assay as described in the instructions for use. 30 NT-proBNP was measured with the proBNP II STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The LoD for this assay was 5 ng/L. 31 Creatinine was measured with the Creatinine Jaffe Gen.2 assay on a cobas c 702 analyzer (Roche Diagnostics, Mannheim, Germany).
HsTnT was measured with the Troponin T hs STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The limit of blank (LoB) for this assay was 3 ng/L and the limit of detection (LoD) was 5 ng/L as described in the instructions for use. 29 HsTnI was measured with the STAT High sensitivity Troponin-I assay on an Architect i1000 analyzer (Abbott, Wiesbaden, Germany). The limit of blank (LoB) was 0.7-1.3 ng/L for this assay as described in the instructions for use. 30 NT-proBNP was measured with the proBNP II STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The LoD for this assay was 5 ng/L. 31 Creatinine was measured with the Creatinine Jaffe Gen.2 assay on a cobas c 702 analyzer (Roche Diagnostics, Mannheim, Germany).
4
Maternal Plasma Thyroid Hormone Analysis
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Maternal blood collected in ethylenediamine tetraacetic acid (EDTA) tubes at 17 wk gestation was shipped overnight, unrefrigerated, to a central biospecimen processing lab (Rønningen et al. 2006 (link)). Plasma was separated and stored in cryovials at and was shipped frozen on dry ice to ARUP Laboratories (Salt Lake City, Utah) for analysis of thyroid hormone concentrations. Thyroid stimulating hormone (TSH) was measured using a quantitative chemiluminescent immunoassay on a Roche Cobas e602 blood analyzer. Triiodothyronine (T3) and thyroxine (T4) were also measured using quantitative electrochemiluminescent immunoassays on the Roche Cobas e602 analyzer. Intra- and interassay CVs for T3, T4, and TSH were . We previously established the reliability of MoBa maternal plasma for measurement of thyroid hormone concentrations, considering delays in processing and storage and freeze-thaw cycles (Villanger et al. 2017 (link)).
5
Comprehensive Clinical and Biochemical Evaluation
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Clinical examinations included questionnaires, medical history, anthropometric measurements, and biochemical analysis. During the examination, the physician recorded the medical history (including previous diseases and drug prescriptions) and drinking frequency and amount. The smoking history was also recorded and distinguished as yes or no.
The anthropometric measurements were performed as previously described, including body weight, standing height, waist circumference, and blood pressure [25 (link), 26 (link)]. Weight and height were measured when the patient was wearing light clothing and no shoes. Waist circumference was measured after the patient exhaled, with the tape measure placed between the lowest rib and the upper edge of the iliac crest. Blood pressure was measured after resting for 5 min. Body mass index (BMI) was calculated as the weight (kg) divided by height (m) squared.
Fasting blood samples were taken from the anterior cubital vein and were used for biochemical analysis. Measurements included liver enzymes, blood lipids, glucose, and uric acid. All of the biochemical values were measured by a Hitachi 7600 clinical analyzer (Hitachi, Tokyo, Japan) using standard methods. Serum 25-hydroxyvitamin D levels were measured with the electro-chemiluminescence immunoassay (ECLIA) platform using the Roche cobas e602 analyzer (Roche Diagnostics GmbH, Germany).
The anthropometric measurements were performed as previously described, including body weight, standing height, waist circumference, and blood pressure [25 (link), 26 (link)]. Weight and height were measured when the patient was wearing light clothing and no shoes. Waist circumference was measured after the patient exhaled, with the tape measure placed between the lowest rib and the upper edge of the iliac crest. Blood pressure was measured after resting for 5 min. Body mass index (BMI) was calculated as the weight (kg) divided by height (m) squared.
Fasting blood samples were taken from the anterior cubital vein and were used for biochemical analysis. Measurements included liver enzymes, blood lipids, glucose, and uric acid. All of the biochemical values were measured by a Hitachi 7600 clinical analyzer (Hitachi, Tokyo, Japan) using standard methods. Serum 25-hydroxyvitamin D levels were measured with the electro-chemiluminescence immunoassay (ECLIA) platform using the Roche cobas e602 analyzer (Roche Diagnostics GmbH, Germany).
6
Quantification of SARS-CoV-2 Antibodies
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Blood samples were collected from study participants. SARS-CoV-2 antibodies were measured using the quantitative Elecsys anti-RBD and semi-quantitative Elecsys anti-N (both measuring total immunoglobulin levels) on the Cobas e602 analyzer (Roche Diagnostics, Rotkreuz, Switzerland). Results for the quantitative Elecsys anti-RBD antibodies are reported as concentrations (U/mL), with a manufacturer’s cutoff of more than 0.8 U/mL considered as positive. Results for the Elecsys anti-N antibodies are reported as cutoff index (signal sample/cutoff or signal calibrator), with values more than 1 considered as positive. Quality controls and coefficients of variation for both assays are provided in Table E1 in this article’s Online Repository at www.jacionline.org . The values of anti-RBD antibodies were log10 transformed before analysis.
7
Serum 25(OH)D and VDBP Analysis
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Each of serum sample was aliquoted into two tubes and stored at −80℃ until the analyses of total 25(OH)D and VDBP levels were performed. Serum total 25(OH) D levels were analyzed using the Elecsys® Vitamin D total II assay and Cobas e602 analyzer (Roche Diagnostics, Mannheim, Germany). Bioavailable 25(OH) D concentrations were calculated using equations reported in previous studies and based on albumin level, total 25(OH)D and VDBP derived from the medical records [2 (link)3 (link)]. VDBP concentrations were measured using the Human Vitamin D BP Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA), according to protocol of manufacturer.
8
Biomarker Measurement in Serum Samples
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All samples were obtained by venipuncture into serum monovette® and centrifuged at 2500 ×g at 20°C for 10 minutes. The aliquoted samples were stored at −80°C until analysis. After thawing the samples were mixed gently by inverting and centrifuged with 2500 ×g for 10 minutes.
Galectin-3 was measured with the Galectin-3 assay on an Architect i1000 analyzer (Abbott, Wiesbaden, Germany). The limit of blank for this assay was 0.8 ng/mL as described in the instructions for use [20 ]. NT-proBNP was measured with the proBNP II STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The limit of detection (LoD) for this assay was 5 pg/mL [21 ]. Serum creatinine was measured with the Creatinine Jaffe Gen. 2 assay on a cobas c 702 analyzer (Roche Diagnostics, Mannheim, Germany).
Galectin-3 was measured with the Galectin-3 assay on an Architect i1000 analyzer (Abbott, Wiesbaden, Germany). The limit of blank for this assay was 0.8 ng/mL as described in the instructions for use [20 ]. NT-proBNP was measured with the proBNP II STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The limit of detection (LoD) for this assay was 5 pg/mL [21 ]. Serum creatinine was measured with the Creatinine Jaffe Gen. 2 assay on a cobas c 702 analyzer (Roche Diagnostics, Mannheim, Germany).
9
Serum Biomarker Measurement Protocol
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All biochemical analyses were performed in a single freeze–thaw cycle >3 assay runs using the same lot of reagents in an accredited laboratory (Changi General Hospital, Singapore). Serum NT-proBNP (proBNP II STAT; Roche Diagnostics, Pensberg, Germany) and hsTnT (STAT; Roche Diagnostics, Pensberg, Germany) were assayed using electrochemiluminescence immunoassay on the Cobas E602 analyzer (Roche Diagnostics Asia-Pacific, Singapore). Serum hsTnI (ARCHITECT STAT High-Sensitive Troponin-I; Abbott Diagnostics, Abbott Park, IL) and galectin-3 (ARCHITECT Galectin-3; Abbott Diagnostics) were determined using chemiluminescent microparticle immunoassay on the ARCHITECT i2000SR analyzer (Abbott Laboratories [Diagnostics], Singapore). Blood samples were collected on the day of CMR.
The manufacturer-reported lower limit of detection (LOD) and the 99th percentile upper reference limit for NT-proBNP were 5 and 135 pg/mL, respectively.31 The LOD and 99th percentile upper reference limit for hsTnT were 5 and 14 pg/mL, respectively.32 The LOD for hsTnI was 1.1 ng/L. We have determined previously that the 99th percentile upper reference limit for hsTnI was 26 ng/L.33 (link) Serum galectin-3 had a manufacturer-reported LOD of 1.0 ng/mL.34 All biochemical concentrations lower than the detection levels were assigned a value equivalent to half the LOD.
The manufacturer-reported lower limit of detection (LOD) and the 99th percentile upper reference limit for NT-proBNP were 5 and 135 pg/mL, respectively.31 The LOD and 99th percentile upper reference limit for hsTnT were 5 and 14 pg/mL, respectively.32 The LOD for hsTnI was 1.1 ng/L. We have determined previously that the 99th percentile upper reference limit for hsTnI was 26 ng/L.33 (link) Serum galectin-3 had a manufacturer-reported LOD of 1.0 ng/mL.34 All biochemical concentrations lower than the detection levels were assigned a value equivalent to half the LOD.
10
Biomarker Analysis in Gallbladder Cancer
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The Cobas E-602 analyzer (Roche, Basel, Switzerland) was used to analyze the levels of serum CEA, CA125 and CA19-9 in 98 GBC patients and 60 healthy volunteers based on electrochemiluminescence (ECL).
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