Coagulation testing included determination of VWF:GPIbM, VWF:Ag, FVIII:C, VWF:CBA and multimers analysis. Both VWF:GPIbM and VWF:Ag were determined with commercially available automated immunoturbidimetric assays, while FVIII:C was measured with a one-stage clotting method, on Atellica COAG 360 analyzer (Siemens Healthcare, Marburg, Germany) following original manufacturer's protocols. VWF:CBA reflecting the ability of binding collagen type 3 was determined by using the enzyme-linked immunosorbent assay Technozym VWF:CBA (Technoclone, Vienna, Austria). VWF multimers were analyzed with agarose gel electrophoresis with direct immunofixation with the Hydragel 5 von Willebrand multimers kit on Hydrasys 2 Scan instrument (Sebia, Lisses, France).
Atellica coag 360 analyzer
Manufactured by Siemens
Sourced in Germany
The Atellica COAG 360 analyzer is a laboratory equipment product designed for coagulation testing. It is a fully automated system that performs a range of coagulation assays to support clinical decision-making. The core function of the Atellica COAG 360 is to provide accurate and reliable coagulation test results.
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4 protocols using atellica coag 360 analyzer
1
Comprehensive Coagulation Profiling Protocol
2
COVID-19 Diagnostic Biomarker Assessment
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The diagnosis of COVID-19 was confirmed in all patients with viral reverse transcriptase PCR using the Xpert® Xpress SARS-CoV-2 (singleplex) cartridge and device (GeneXpert, Cepheid GmbH, 47807 Krefeld, Germany), as previously described [33 (link)]. Interleukin-6 (IL-6), ferritin, NTproBNP, cTnT, and C-reactive protein (CRP) were measured with commercial immunoassays on a COBAS 8000 analyzer (Roche Diagnostics, Rotkreuz, Switzerland). We measured vitamin K by a newly developed LC-MS/MS method [32 (link)]. The overall precision of all analytes (combination of intra and inter-assay CVs) ranged between 4.8% and 17.7%. D-Dimer, prothrombin time, activated partial thromboplastin time (aPTT), antithrombin (AT), protein C, free protein S, and factor XIII were analyzed on a Siemens Atellica COAG-360 analyzer (Siemens Healthineers, Marburg, Germany).
3
Siemens INNOVANCE Heparin anti-Xa Assay Protocol
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The Siemens INNOVANCE® Heparin anti-Xa assay was selected for determining the anti-Xa activity. This one-stage chromogenic assay is designed to measure the drug level of unfractionated heparin (UFH) as well as low molecular weight heparin (LMWH) (INNOVANCE® Heparin anti-Xa assay application sheet; Siemens Healthineers, Marburg, Germany). The reagent contains dextran sulfate but not exogenous antithrombin. The instructions provided by the manufacturer were strictly followed. In brief, a five-step calibration curve was applied using the calibrators provided by the manufacturer (0.0, 0.42, 0.86, 1.26, and 1.67 U/ml). Samples were rapidly thawed and gently mixed at 37°C. Patient plasma was pre-diluted (1:2) and added to the reagent containing coagulation factor Xa and a chromogenic substrate. The formation of paranitroaniline was quantified optically at a wavelength of 405 nm. The measurements were conducted in batches on an Atellica COAG 360 analyzer (Siemens Healthineers, Marburg, Germany) (23 (link)). Measurements were performed blinded to the LC-MS/MS test results.
4
Chromogenic Assays for Factor VIII and IX
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Reagents and calibrators were used on the Atellica COAG 360 analyzer (Siemens Healthineers) according to the instructions provided by the manufacturer (Table 1 ). Factor VIII chromogenic activity was measured using two different CSA methods; CSA‐1 (see below) and CSA‐2 (Siemens). Methods not provided by Siemens and thus required LDT were chromogenic assays for FVIII (CSA‐1, Chromogenix, IL Company) and FIX (Rossix), with normal reference plasma (NRP) from Precision Biologic as calibrator. Calibration included a high (calibration points ≥ 0.125 IU/ml, for samples > 0.20 IU/ml) and a low (calibration points ≤ 0.25 IU/ml, for samples ≤ 0.20 IU/ml) calibration curves using at least 5 points per curve. Reagent for prothrombin time using Owren was from Medirox with the national INR calibrator from Equalis. Controls were from Siemens, except for the low FIX controls (0.1 and <0.05 IU/ml), which were from Precision Biologic. The same reagents were used on the BCS‐XP (Siemens) except for STA‐PTT automate (Diagnostica Stago) and Thromborel S (Siemens) as OSA reagents and Coamatic heparin (Chromogenix) as anti‐FXa activity (LMWH) reagent.
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