Blood samples were drawn at 8 a.m. after an overnight fast. Samples were centrifuged and sera kept frozen at − 20 °C until analysis. Analysis of serum TSH was performed with CLIA with the aid of an Abbott ARCHITECT instrument (Abbott Diagnostics Division). Total coefficient of variation (CV) was < 3.3%, functional sensitivity was 0.0038μUI/mL; reference range [99% confidence interval (CI)]: 0.35–4.94μUI/mL; fT4 was measured by Abbott ARCHITECT instrument (Abbott Diagnostics Division; total CV was < 7%, sensitivity of the assay was < 0.4 ng/dL; reference range (99% CI): 0.70–1.48 ng/dL (conversion factor ng/dL *12.87 = pmol/L). TPOAb were measured by an Abbott ARCHITECT instrument (Abbott Diagnostics Division). Total CV was < 7.6%, sensitivity of the assay was 0.16 IU/mL; reference range: < 5.61 IU/mL.
Architect instrument
Manufactured by Abbott
Sourced in Germany, United States
The Architect instrument is a clinical chemistry and immunoassay analyzer developed by Abbott. Its core function is to automate the analysis of various biological samples, such as blood, urine, and body fluids, to provide healthcare professionals with accurate and reliable test results for diagnostic purposes.
Automatically generated - may contain errors
Lab products found in correlation
Architect analyser, by Abbott (1 mentions)
Anti hbe, by Roche (1 mentions)
Bigdye terminator v3.0 cycle sequencing ready reaction kit, by Thermo Fisher Scientific (1 mentions)
Abi 3130xl genetic analyzer, by Thermo Fisher Scientific (1 mentions)
Modular p800 analyzer, by Roche (1 mentions)
Imx instrument, by Abbott (1 mentions)
Sars cov 2 igg 2 quant, by Abbott (1 mentions)
Bep 3 system, by Siemens (1 mentions)
Cobas instrument, by Roche (1 mentions)
Liaison instrument, by DiaSorin (1 mentions)
Lightcycler multiplex rna virus master, by Roche (1 mentions)
Lightmix modular eav rna extraction control, by TIB Molbiol (1 mentions)
Lightcycler 480 instrument 2, by Roche (1 mentions)
10 protocols using architect instrument
1
Serum TSH, fT4, and TPOAb Analysis
2
SARS-CoV-2 Antibody Detection Assays
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Aliquots from all patient samples presented in this work underwent SARS-CoV-2 diagnostic testing on an Architect instrument (Abbott). The SARS-CoV-2 IgM, and the indirect format SARS-CoV-2 IgG II Quant Architect assays were performed using commercially available assay kits, according to the package instructions (link to the package insert: https://www.corelaboratory.abbott/us/en/offerings/segments/infectious-disease ). The direct format SARS-CoV-2 Total Ig (S) research assay was developed in-house (Abbott) to detect antibodies against RBD of the S1 subunit of the spike protein of SARS-CoV-2 virus in patient plasma. In this one-step assay, patient sample (35 µL), SARS-CoV-2 spike protein-coated magnetic microparticles (50 µL, 0·075% solids) and acridinium-labelled spike protein conjugate (50 µL, 80 ng/mL) were combined, incubated, washed, and measured on an Abbott Architect analyser.
3
Hepatitis B Serological Profiling and Genotyping
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HBsAg and anti-HBc were run on the Architect instrument (Abbott, Germany) and values equal to or greater than 1.00 Sample/Cutoff (S/CO) were considered positive. Anti-HBs titres (Architect, Abbott, Germany) were reported as milli-international units per ml (mIU/ml) according to WHO international reference standard (Hollinger and Dreesman, 1986). Samples equal to or greater than 10mIU/ml were considered positive for anti-HBs. Samples positive for HBsAg were tested for HBeAg and anti-HBe (Architect, Abbot, Germany) with cutoff at 1.00 S/CO.
Hepatitis B viral load was performed on HBsAg positive samples using the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0, quantitative assay (Roche Molecular Systems, New Jersey, USA). Sequencing was performed on the polymerase region (nucleotides 2624–1240), overlapping the complete S region (nucleotides 2848–2835) using BigDye Terminator v3.0 Cycle Sequencing Ready Reaction Kit (Applied Biosystems, Foster City, CA, USA) on the ABI 3130XL Genetic Analyzer (Applied Biosystems) [38 (link)]. Neighbour-joining phylogenetic analysis with a bootstrap of 1000 replicates was conducted using Molecular Evolutionary Genetics Analysis (MEGA 5)[39 (link)].
Hepatitis B viral load was performed on HBsAg positive samples using the COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0, quantitative assay (Roche Molecular Systems, New Jersey, USA). Sequencing was performed on the polymerase region (nucleotides 2624–1240), overlapping the complete S region (nucleotides 2848–2835) using BigDye Terminator v3.0 Cycle Sequencing Ready Reaction Kit (Applied Biosystems, Foster City, CA, USA) on the ABI 3130XL Genetic Analyzer (Applied Biosystems) [38 (link)]. Neighbour-joining phylogenetic analysis with a bootstrap of 1000 replicates was conducted using Molecular Evolutionary Genetics Analysis (MEGA 5)[39 (link)].
4
Therapeutic Drug Monitoring Assays
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Whole blood Tac concentrations were measured using the CMIA (chemiluminescent microparticle immunoassay) on the Architect instrument (Abbott Laboratories, Lake Forest, IL, USA) and CsA concentrations using the CEDIA PLUS assay (Cloned Enzyme Donor Immunoassay; Microgenics Corporation, Fremont, CA, USA) on a Modular P800 analyzer (Roche Diagnostics, Rotkreuz, Switzerland).
5
Tacrolimus Concentration Measurement Techniques
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In the Brisbane dataset, all tacrolimus concentration measurements were made using liquid chromatography-tandem mass spectrometry assay (LC-MS/MS) 24 . In the Oslo dataset, 80% of concentrations were measured with chemiluminescent microparticle immunoassay (CMIA, analyzed on the Architect® instrument, Abbott Laboratories, Abbott Park, IL 25 (link)), 11% with LC-MS/MS 26 (link) and 9% with microparticle enzyme immunoassay (MEIA, analyzed on the IMx® instrument, Abbott Laboratories 27 (link)). In the external evaluation dataset, all concentrations were measured with CMIA. Concentrations (C) measured with CMIA and MEIA were converted to corresponding LC-MS/MS equivalents using an equation derived from linear regression as described previously (Equation 1 ) 7 (link):
The LC-MS/MS assay used in Brisbane was linear over the range between 0.5 to 50 μg l−1. The imprecision coefficient of variation (CV) was 5%. The LC-MS/MS assay used in Oslo had a lower limit of quantification (LLOQ) of 1.1 μg l−1 and a CV of 5.2%. The CMIA had a LLOQ of 1.0 μg l−1 and CVs of 9% at 2.3 μg l−1 and 6% at 7.0 μg l−1. The MEIA had a LLOQ of 3.0 μg l−1 and CVs of 13% at 5 μg l−1 and 7% at 23 μg l−1.
The LC-MS/MS assay used in Brisbane was linear over the range between 0.5 to 50 μg l−1. The imprecision coefficient of variation (CV) was 5%. The LC-MS/MS assay used in Oslo had a lower limit of quantification (LLOQ) of 1.1 μg l−1 and a CV of 5.2%. The CMIA had a LLOQ of 1.0 μg l−1 and CVs of 9% at 2.3 μg l−1 and 6% at 7.0 μg l−1. The MEIA had a LLOQ of 3.0 μg l−1 and CVs of 13% at 5 μg l−1 and 7% at 23 μg l−1.
6
Quantifying Spike Protein Antibodies Post-BNT162b2
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Blood samples were collected once from the second to the sixth week after the second BNT162b2 dose (end of study). Blood was collected in serum tubes without anticoagulants, left to coagulate at room temperature for 30 minutes, and centrifuged at 4 °C 1800xG for 10 min. Sera were transferred to clean vials and stored at -20 °C pending analysis. Samples were analyzed with SARS-CoV-2 IgG II Quant (Abbott) on the Architect instrument (Abbott) to quantitatively and qualitatively detect IgG antibodies against the Spike protein S1 receptor-binding domain (RBD). Antibody Units per ml were converted into WHO binding antibody units (BAU/ml), according to the manufacturer's indications. A threshold of at least 7.1 BAU/ml was the cut-off for a positive response to immunization. Samples were also tested qualitatively for the presence of IgGs against the nucleocapsid antigen (anti-N) to verify a possible previous asymptomatic SARS-CoV-2 infection.
7
Automated CMV Serological Testing
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The CMV serological testing of blood donors was performed using the fully automated CMV immunoglobulin M (IgM) and IgG tests on the Architect instrument (Abbott Laboratories, Abbott Park, IL) or the BEP® III system (Siemens Healthcare) by the diagnostics department of the Institute for Medical Microbiology and Hygiene (University of Regensburg, Germany). CMV IgG-serology was used as primary reference measurement procedure (gold standard method).
8
Tacrolimus Whole-Blood Concentration Assay
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All blood samples for tacrolimus whole-blood concentration measurement were drawn immediately before the morning dose. Concentrations were determined using the chemiluminescent microparticle immunoassay (CMIA, analysed on the Architect Instrument; Abbott Laboratories, Abbot Park, IL). 21 The assay was consistently applied throughout the study period. The lower limit of quantification was 1.0 µg/L.
The coefficients of variation of the between series imprecision were 6% at 2 µg/L and 3.5% at 7.2 µg/L, respectively.
The coefficients of variation of the between series imprecision were 6% at 2 µg/L and 3.5% at 7.2 µg/L, respectively.
9
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).
10
SARS-CoV-2 Detection by RT-PCR and Serological Testing
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For the detection of viral RNA by RT-PCR, each tube sample contained a nasopharyngeal and an oropharyngeal swab immersed in virus preservation solution. RNA was extracted with the Qiacube extractor by using the spin-column Qiamp virus minikit (Qiagen, Hilden, Germany). The reported RT-PCR results for the SARS-CoV-2 E gene were obtained with the LightCycler® Multiplex RNA Virus Master (Roche Life Science, Penzberg, Germany) at a LightCycler® 480 Instrument II (Roche Life Science, Penzberg, Germany), and it included a RNA extraction control (LightMix® Modular EAV RNA Extraction Control; Tib Molbiol, Berlin, Germany). Positive and negative controls were routinely included with each batch of tests. Relative quantification of the sample crossing points (cp) was automatically inferred with the LightCycler software.
The serological test used was a chemiluminescent microparticle immunoassay for the qualitative detection of IgG against SARS-CoV-2 nucleoprotein (Abbott Diagnostics, Chicago, IL, USA). Serum samples were run on the Abbott Architect instrument following the manufacturer’s instructions. The amount of IgG antibodies to SARS-CoV-2 in each sample was determined by comparing its chemiluminescent relative light unit (RLU) to the calibrator RLU (index S/C). A signal/cut-off (S/CO) ratio of ≥1.4 was interpreted as reactive and an S/CO ratio of <1.4 was interpreted as non-reactive.
The serological test used was a chemiluminescent microparticle immunoassay for the qualitative detection of IgG against SARS-CoV-2 nucleoprotein (Abbott Diagnostics, Chicago, IL, USA). Serum samples were run on the Abbott Architect instrument following the manufacturer’s instructions. The amount of IgG antibodies to SARS-CoV-2 in each sample was determined by comparing its chemiluminescent relative light unit (RLU) to the calibrator RLU (index S/C). A signal/cut-off (S/CO) ratio of ≥1.4 was interpreted as reactive and an S/CO ratio of <1.4 was interpreted as non-reactive.
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