Thrombin reagent

Manufactured by Siemens

Sourced in Germany

The Thrombin Reagent is a laboratory product manufactured by Siemens. It is a liquid reagent used in coagulation testing procedures. The primary function of the Thrombin Reagent is to initiate the clotting of blood samples.

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Lab products found in correlation

Cs2100i, by Sysmex (4 mentions) Thromboclotin, by Siemens (2 mentions) Vacutainer sst 2 advance tube, by BD (1 mentions) Zymutest, by Hyphen Biomed (1 mentions) Sta r, by Stago (1 mentions) Coulter act diff analyzer, by Beckman Coulter (1 mentions) Trisodium citrate tubes, by BD (1 mentions) D dimer, by Siemens (1 mentions) Vacutainer system, by BD (1 mentions) Actine fsl, by Siemens (1 mentions) Innovin pt, by Siemens (1 mentions) Modular p800 analyzer, by Roche (1 mentions) Architect ci8200, by Abbott (1 mentions) Actin fs, by Siemens (1 mentions)

10 protocols using thrombin reagent

Biomarkers for Pulmonary Fibrosis in COVID-19

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We retrospectively extracted from the medical file levels of KL-6 as marker of pulmonary fibrosis). Fibrinogen (a coagulopathy marker [19 (link), 20 (link)]), lactate dehydrogenase (LDH) (a lung injury marker [18 (link), 19 (link), 21 (link)]), C-reactive protein (CRP) (an inflammatory marker) and platelet/lymphocytes ratio (PLR) (a promising prognostic marker of COVID-19 severity [22 (link)]) were also assessed. Pulse oximetry (SPO₂) was monitored during patients’ hospitalization to assess the respiratory status on day 7 post-SARS-CoV-2 detection. Respiratory insufficiency was defined as lower than 90%. Blood samples were collected in BD Vacutainer SST II Advance tubes and centrifuged to quantify LDH, CRP, and KL-6. The reference values for LDH and CRP on serum are respectively between 125–220 U/L and 0–5 mg/L. For KL-6 measurement, we used a chemiluminescent light immunoassay on the Fujirebio Lumipulse G1200 instrument (Tokyo, Japan). The threshold for high levels of KL-6 was 453.5 U/ml. Other analyses were run on the Abbott Alinity platform (Abbott Park, IL, USA). Blood samples were collected in citrate tubes for fibrinogen measurement, and a coagulometric test was made using Thrombin reagent (Siemens, Germany). The reference values for KL-6 on serum are between 118–627 U/ml.

Cambier M., Henket M., Frix A.N., Gofflot S., Thys M., Tomasetti S., Peired A., Struman I., Rousseau A.F., Misset B., Darcis G., Moutschen M., Louis R., Njock M.S., Cavalier E, & Guiot J. (2022). Increased KL-6 levels in moderate to severe COVID-19 infection. PLOS ONE, 17(11), e0273107.

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Comprehensive Coagulation Factor Analysis

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FII, FV, FVII, FX (Neoplastine R), FVIII, FIX, FXI, FXII (CK Prest) activity levels were measured by clotting assays using specific factor‐depleted plasma as substrate on Sta‐R (Stago, Paris, France). Fibrinogen level (Clauss method, Thrombin Reagent; Siemens, Marburg, Germany), FXIII activity and vWF antigen and activity (FXIIIact/subs, vWF Reag and vWF Ac Reagens; Siemens) measurements were performed on a Sysmex CS2100i. α2‐antiplasmin level was measured using a chromogenic assay (Stachrom; Stago), tissue plasminogen activator (tPA) and plasminogen activator inhibitor 1 (PAI‐1) activity were measured using bio‐immunoassays (Zymutest, Hyphen‐BioMed, Neuville‐sur‐Oise, France).

Vries M.J., van der Meijden P.E., Kuiper G.J., Nelemans P.J., Wetzels R.J., van Oerle R.G., Lancé M.D., ten Cate H, & Henskens Y.M. (2018). Preoperative screening for bleeding disorders: A comprehensive laboratory assessment of clinical practice. Research and Practice in Thrombosis and Haemostasis, 2(4), 767-777.

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Coagulation Screening for Lupus Anticoagulant

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Factor deficiencies and undisclosed anticoagulation were excluded by performing coagulation screening tests prior to the LA assays. APTT, prothrombin time (PT), thrombin time, and fibrinogen for Clauss method tests were measured by a Sysmex CS‐2100i (Sysmex UK) using Actin FS, Dade Innovin, Thromboclotin, and Thrombin‐Reagent (Siemens Healthineers, Marburg, Germany), respectively. As a LA‐insensitive routine APTT reagent, Actin FS was employed and thus the reagent is suitable for exclusion of other cause of clotting time prolongation.8, 16

Kumano O, & Moore G.W. (2017). Lupus anticoagulant mixing tests for multiple reagents are more sensitive if interpreted with a mixing test‐specific cut‐off than index of circulating anticoagulant. Research and Practice in Thrombosis and Haemostasis, 2(1), 105-113.

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Evaluating Clot Analogue Mechanics

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Donor age and sex was recorded. RBCs, platelets and white blood cells were counted with a Coulter Counter (COULTER® AC•T diff™ Analyzer, Beckman Coulter, CA) (Fig. 1e). Fibrinogen levels were measured using the Clauss assay (Thrombin Reagent, Siemens Healthineers, Erlangen, Germany) on the Sysmex CS5100 coagulation analyzer (Siemens Healthineers, Diagnostics B.V., Newark, DE, USA) (Fig. 1f). Since contraction has been shown to alter clot analogue mechanics, the degree of clot contraction was assessed gravimetrically as the percentage weight of the expelled serum compared with the weight of the total blood mixture (Fig. 1g).^{13 (link)}

Cahalane R.M., de Vries J.J., de Maat M.P., van Gaalen K., van Beusekom H.M., van der Lugt A., Fereidoonnezhad B., Akyildiz A.C, & Gijsen F.J. (2023). Tensile and Compressive Mechanical Behaviour of Human Blood Clot Analogues. Annals of Biomedical Engineering, 51(8), 1759-1768.

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Coagulation Parameters Measurement Protocol

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Blood from all subjects was collected into 3.2% trisodium citrate tubes (9:1 v/v, Becton Dickinson) and centrifuged 15 min at 2500g at room temperature, followed by 20 000g for 10 min. The obtained platelet‐poor plasma was stored in aliquots at −80°C until thawed at 37°C immediately before use. Fibrinogen levels were measured using the Clauss assay (Thrombin Reagent, Siemens Healthineers), factor VIII (FVIII) levels using a one‐stage clotting assay and FXIII levels with a chromogenic assay using the Berichrom FXIII kit (Siemens Healthineers). In addition, levels of D‐dimer (Siemens Healthineers) and anti‐Xa (Werfen) were routinely measured. All parameters were measured on the Sysmex CS5100 coagulation analyzer (Siemens Healthcare Diagnostics B.V.). Other relevant parameters (e.g., C‐reactive protein) were routinely measured. PAI‐1 levels were measured using the human Serpin E1/PAI‐1 DuoSet ELISA kit (DY1786, R&D Systems) according to the protocol provided by the manufacturers, with some minor changes in the volumes of samples, detection antibody, and streptavidin‐horseradish peroxidase (50 µl instead of 100 μl). Values above the standard of the ELISA were given the maximum value of the standard (40 ng/ml).

de Vries J.J., Visser C., Geers L., Slotman J.A., van Kleef N.D., Maas C., Bax H.I., Miedema J.R., van Gorp E.C., Goeijenbier M., van den Akker J.P., Endeman H., Rijken D.C., Kruip M.J, & de Maat M.P. (2022). Altered fibrin network structure and fibrinolysis in intensive care unit patients with COVID‐19, not entirely explaining the increased risk of thrombosis. Journal of Thrombosis and Haemostasis, 20(6), 1412-1420.

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Thrombin-Induced Clot Dissolution

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The ELT was performed similar to Kowalski et al.13 Differently, the precipitate was dissolved in 1 mL Veronal buffer and the reaction was started by adding 25 μL thrombin 100 U/mL (Thrombin Reagent; Siemens). Every 10 to 15 minutes, clot resolution was assessed visually, and every 5 minutes when clot dissolution was almost reached.

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Plasma Fibrinogen Measurement Protocol

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Blood samples were acquired at least three months after delivery. Blood sampling was performed by venipuncture using the Vacutainer system (Becton Dickinson, Utrecht, The Netherlands) containing 3.2% sodium citrate. Citrated blood was centrifuged two times at 2000× g for 10 min at room temperature, followed by 14,000× g for 10 min centrifugation of plasma at room temperature. Plasma samples were stored in aliquots at −80 °C until analysis. Fibrinogen activity according to the Clauss method (Thrombin Reagent, Siemens Healthcare Diagnostics B.V., Breda, The Netherlands) was measured on a Sysmex CS5100 (Siemens Healthcare Diagnostics B.V.).

Daraei A., Pieters M., Baker S.R., de Lange-Loots Z., Siniarski A., Litvinov R.I., Veen C.S., de Maat M.P., Weisel J.W., Ariëns R.A, & Guthold M. (2021). Automated Fiber Diameter and Porosity Measurements of Plasma Clots in Scanning Electron Microscopy Images. Biomolecules, 11(10), 1536.

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Coagulation Biomarkers Analysis

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Fibrinogen level (Clauss method, Thrombin reagent, Siemens), PT (Innovin Pt, Siemens) and activated partial thromboplastin time (aPTT; Actine FSL, Siemens) were determined in PPP on a Sysmex CS2100i.

Hulshof A.M., Olie R.H., Vries M.J., Verhezen P.W., van der Meijden P.E., ten Cate H, & Henskens Y.M. (2021). Rotational Thromboelastometry in High-Risk Patients on Dual Antithrombotic Therapy After Percutaneous Coronary Intervention. Frontiers in Cardiovascular Medicine, 8, 788137.

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Immunocomplex-Based Protein Quantification Protocol

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Samples were processed as described above, concentrations of ceruloplasmin and serum albumin were determined nephelometrically on a BN-Prospec (Siemens, Tarrytown, NY) after immuno-complexation with their respective antisera (Siemens). Concentrations of haptoglobin, immunoglobulins alpha, gamma and mu as well as serotransferrin were determined by turbidity measurements on a Modular P800 analyzer (Roche, Basel, Switzerland) following immuno-complexation with their respective Tina-Quant antisera (Roche). After immuno-complexation with their respective antisera (Abbott, Chicago, IL), concentrations of Complement C3, C4 and apolipoproteins A1 and B-100 were determined by turbidity measurements on an ARCHITECT ci8200 (Abbott). Fibrinogen concentration was determined by measuring plasma clotting using a thrombin reagent (Siemens) on a Sysmex CA-7000 (Siemens). Reference and assay ranges are given in Table 1 for the different assays employed.

Kramer G., Woolerton Y., van Straalen J.P., Vissers J.P., Dekker N., Langridge J.I., Beynon R.J., Speijer D., Sturk A, & Aerts J.M. (2015). Accuracy and Reproducibility in Quantification of Plasma Protein Concentrations by Mass Spectrometry without the Use of Isotopic Standards. PLoS ONE, 10(10), e0140097.

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Coagulation Screening Panel Assessment

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Prothrombin time (PT), APTT, fibrinogen by the Clauss method, and thrombin time were performed as coagulation screening tests. The reagents were Dade Innovin, Actin FS, Thrombin‐Reagent, and Thromboclotin (Siemens Healthineers), respectively. All tests were performed in a Sysmex CS‐2100i (Sysmex UK, Milton Keynes, UK).

Kumano O, & Moore G.W. (2019). Ruling out lupus anticoagulants with mixing test–specific cutoff assessment and the index of circulating anticoagulant. Research and Practice in Thrombosis and Haemostasis, 3(4), 695-703.

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