Innovance d dimer assay

After the measurement of pancreatic blood flow, blood samples were taken from the abdominal aorta. The prothrombin time measured as international normalized ratio (INR) was determined in fresh blood, using Alere INRatio^® 2 PT/INR Monitoring Systems and Alere INRatio^® PT/INR Monitoring System Test Strips (Alere San Diego, Inc, San Diego, CA, USA).
Plasma D-Dimer concentration was determined using an immunoturbidimetric assay (Innovance D-Dimer Assay, Siemens Healthcare GmbH, Marburg, Germany) on automatic coagulation analyzer BCS XP System (Siemens Healthcare Diagnostics, Erlangen, Germany).
Serum lipase and amylase activity was determined with a Kodak Ectachem DT II System analyzer (Eastman Kodak Company, Rochester, NY, USA) using Lipa and Amyl DT Slides (Vitros DT Chemistry System, Johnson & Johnson Clinical Diagnostic, Inc., Rochester, NY, USA).
Serum concentration of interleukin-1β (IL-1β) was measured using the Rat IL-1 beta ELISA Kit (Biorbyt Ltd., Cambridge, UK).

Conventional coagulation tests were performed in all study participants. Complete blood counts were measured on a Sysmex XE-2100 analyzer (Roche, Chicago, IL, USA). aPTT, PT, INR, fibrinogen and D-dimers were determined on a BCS^® XP System Hemostasis analyzer (Siemens Healthcare Diagnostics, Marburg, Germany).More specifically, Pathromtin SL (Siemens Healthcare Diagnostics, Marburg, Germany) was used to assess aPTT and Thromborel S Reagent for PT calculation. Determination of fibrinogen concentrations were made performing a modification of the Clauss method with Fibrinogen Multifibren U reagent (Siemens Healthcare Diagnostics, Marburg, Germany). D-dimers were assessed by the INNOVANCE D-Dimer assay (Siemens Healthcare Diagnostics, Marburg, Germany), a particle-enhanced immunoturbidimetric method.

Demographic data, prior history, clinical, physical examinations, laboratory tests and radiological examinations were obtained from electronic medical records. D-dimer was measured by the Sysmex CA7000 analyzers using the INNOVANCE D-dimer assay (Siemens Healthcare Diagnostics Products GmbH, Marburg, Germany). Liver cirrhosis was diagnosed by radiological evidence of liver nodularity or endoscopic signs of portal hypertension at or before admission. Survival rates were obtained through patient medical records or by direct contact with patients or their kins. The model for end-stage liver disease (MELD) [14 (link)], MELD sodium (MELD-Na) [15 (link)] scores, CLIF Consortium ACLF score (CLIF-C ACLFs) [16 (link)] and CLIF Consortium Acute Decompensation score (CLIF-C ADs) [17 (link)] were calculated. Organ failures were determined according to the CLIF Consortium Organ Failure score (CLIF-C OFs) [16 (link)]. Diagnosis of European Association for the Study of liver (EASL)-ACLF were according to CANONIC Study [1 (link)].

Venous blood samples were drawn with minimal stasis using atraumatic venipuncture at 08.00–11.0 AM, after an overnight fast. All measurements were performed in VTE patients after 3 months of anticoagulant therapy since the index event. Patients were drawn >24 hours since the last dose of NOACs. Complete blood count, glucose, creatinine, lipid profiles, and INR were assayed by routine laboratory techniques. High-sensitivity C-reactive protein (hs-CRP) was determined by immunoturbidimetry (Roche Diagnostics GmbH, Mannheim, Germany). Plasma D-dimer was measured with the Innovance D-dimer assay (Siemens, Marburg, Germany). Plasma α₂-antiplasmin and plasminogen were measured by chromogenic assays (STA Stachrom antiplasmin and Stachrom plasminogen, Diagnostica Stago, Asniéres, France). Plasma PAI-1 antigen was measured by an ELISA kit (Hyphen). For evaluation of clot properties and thrombin generation, venous blood samples were mixed with 3.2% trisodium citrate (vol/vol, 9 : 1), then centrifuged at 2000 × g for 10 min within 30 minutes of the draw, and stored in aliquots at -80°C until analysis. All measurements were performed by technicians blinded to the origin of the samples. Intra-assay and interassay coefficients of variation were 5-7%.

Venous blood samples were collected at baseline from all participants using a 21-gauge needle. The blood was drawn into 3.5-mL VACUETTE 9NC coagulation 3.2% trisodium citrate (0.109 mol/L) tubes (Greiner Bio-One). All tubes were kept in a vertical position with no agitation during transportation to the laboratory. The samples were processed within 2 hours of extraction. Platelet-poor plasma (PPP) was obtained by centrifugation, as our group described previously.^{19 (link)} In brief, the samples underwent centrifugation at 1500g for 30 minutes at 4 °C, with no brake application. The resulting PPP was then collected and stored at –80 °C for future use. The levels of D-dimer and sP-selectin in the PPP of both the case and control groups were measured using commercially available kits. The INNOVANCE D-Dimer Assay (Siemens Healthineers) was used to measure D-dimer, with a lower detection limit of 0.19 mg/L fibrinogen-equivalent units. For sP-selectin, the Human sP-selectin/CD62P Immunoassay (R&D Systems) was used according to the manufacturer’s guidelines. The results were expressed as µg/L for D-dimer and ng/mL for sP-selectin, respectively. The assessment of blood cell count, platelets, and hemoglobin levels was performed using standardized clinical laboratory procedures with a Sysmex XN-10 Automated Hematology Analyzer (Sysmex).

Data were abstracted from the complete electronic medical chart (EMR), including sociodemographic characteristics, reproductive history, gynecologic history, previous contraceptive use, obstetric characteristics, and the results of biomarkers and imaging examinations. At 42 days postpartum, all women came to the hospital for regular examination and these data were also collected.
The data of biomarkers containing D-dimer level, white blood cell counts, platelet counts, fibrinogen level, brain natriuretic peptide (BNP), and other plasma lipid levels were collected. Blood samples for testing D-dimer were collected on postpartum day 2, and samples for other biomarkers were collected on postpartum day 1. All hematology samples were sent immediately to the laboratory for testing after collection. Plasma concentrations of D-dimer was measured by using an immunoturbidimetric Innovance D-Dimer Assay (Siemens Healthcare Diagnostics, Marburg, Germany) and a Sysmex CN-6000 instrument (TOA Medical electronics Co., Kobe, Japan). All biomarker values were obtained from the same laboratory affiliated with the hospital.