Blood samples were collected at prevaccination and 43 days after doses 1 and 2. Antibody titres were measured using a commercial ELISA (Enzygnost, Dade Behring, Marburg, Germany) with cut-off values of 150 mIU/mL (measles), 231 U/mL (mumps) and 4 IU/mL (rubella). For varicella, antibody titres were measured using an immunofluorescence assay (Virgo, Hemagen Diagnostics, Columbia, Maryland, USA; assay cut-off value of 4/dilution).
Enzygnost
Manufactured by Siemens
Sourced in Germany, Canada
Enzygnost is a laboratory equipment product manufactured by Siemens. It is designed for the detection and quantification of specific enzymes in biological samples. The core function of Enzygnost is to provide accurate and reliable analytical results for researchers and healthcare professionals.
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39 protocols using enzygnost
1
Antibody Titers After Vaccination
2
Alcohol Consumption and Liver Disease
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Subjects who never drank alcohol were classified as “non-drinkers”; those who reported drinking alcohol often were classified as “habitual drinkers”. Fatty liver disease, cirrhosis, gallstones, and splenomegaly were all diagnosed by abdominal sonography[6 (link)]. Fatty liver disease was diagnosed if the liver had homogenously increased echogenicity and a smooth surface. Cirrhosis was diagnosed if there was increased parenchymal echogenicity, poor tissue penetration, and parenchymal inhomogeneity. Liver cysts and hemangiomas were diagnosed based on their characteristic sonographic signals. Gallstones were diagnosed based on a characteristic increased echogenicity anywhere in the biliary tree, and included patients who had “silent stones”. Splenomegaly was diagnosed if the spleen was greater than 12 cm in length and had a maximal transverse diameter of 5 cm.
All venous blood samples were obtained in the morning after a 12 h overnight fast. Globulin, albumin, and other serum parameters were analyzed by a biochemical autoanalyser (Hitachi 736-15, Tokyo, Japan) at the Department of Clinical Laboratory within 4 h of collection. Hepatitis B surface antigen was detected by an ELISA test from Enzygnost, Dade Behring GmbH (Marburg, Germany) and the antibody to hepatitis C virus was detected by an ELISA test from Abbott (HCV EIA 3.0, Abbott Laboratories. Abbott Park, Illinois).
All venous blood samples were obtained in the morning after a 12 h overnight fast. Globulin, albumin, and other serum parameters were analyzed by a biochemical autoanalyser (Hitachi 736-15, Tokyo, Japan) at the Department of Clinical Laboratory within 4 h of collection. Hepatitis B surface antigen was detected by an ELISA test from Enzygnost, Dade Behring GmbH (Marburg, Germany) and the antibody to hepatitis C virus was detected by an ELISA test from Abbott (HCV EIA 3.0, Abbott Laboratories. Abbott Park, Illinois).
3
Serological Measurement of MMR Antibodies
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The measurement of MMR IgG antibodies was carried out by means of the commercial enzyme-linked immunosorbent assay (ELISA) Enzygnost (Dade Behring, Marburg, Germany). Antibody levels of Ru and Me were reported as positive (higher than 10 or 350 IU/L, respectively), negative (lower than 5 or 150 IU/L, respectively) or equivocal (5–10 or 150–350 IU/L, respectively). Mu antibody measurement was quantitative. According to the Center for Disease Control and Prevention (CDC) recommendations [5 ], equivocal results were processed as negative.
4
Measles IgG Antibody Quantification
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The measles IgG antibodies were measured by means of commercial enzyme-linked immunosorbent assay (EIA) Enzygnost (Dade Behring, Marburg, Germany). Antibody levels of measles were reported as positive (higher than 350 IU/mL), negative (lower than 150 IU/mL), or equivocal (150–350 IU/mL). Equivocal results had been statistically processed as negative according to CDC recommendations [16 ].
5
Multimodal Hemostasis Assessment in Liver Injury
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Blood samples for arterial blood gas analysis, whole blood cell count, thromboelastometry and plasma preparation were collected at the following time points: after splenectomy (baseline), at the end of haemodilution, and at 10, 60 and 120 minutes after liver injury. Haemoglobin concentration, pH, partial pressure of oxygen and pCO2 were measured using a blood gas analyser (ABL500, Radiometer, Brønshøj, Denmark). Prothrombin time (PT), activated partial thromboplastin time (aPTT) and fibrinogen concentration were determined by standard laboratory methods using the appropriate kits from Dade Behring (Marburg, Germany) on an MC 4 plus steel-ball coagulometer (Merlin Medical, Lemgo, Germany); the fibrinogen concentration was measured using the Clauss assay. Thrombin–antithrombin complexes (TAT) were quantified by ELISA (Enzygnost, Dade Behring, Marburg, Germany).
6
Serological Detection of Viral Infections
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A capture IgM ELISA assay (Venture Technologies, Sarawak, Malaysia) that utilizes inactivated antigens from JEV and DENV1–DENV4 was used to detect and distinguish between JEV- and DENV-specific IgM antibodies in CSF, and was performed as previously described [23] (link).
A chemiluminescent microparticle immunoassay (CMIA, Abbott diagnostic division, Lake Forest, Illinois 60045, USA) was used for the detection of IgM antibodies to rubella virus in serum, and was performed according to the manufacturer's instructions.
For detection of EBV antibodies in serum, an IgG/IgM ELISA (Enzygnost, Dade-Behring, Marburg, Germany) using recombinant viral capsid antigen (VCA) and EBV nuclear antigen (EBNA) was used, and was performed according to the manufacturer's instructions.
Detection of CMV-specific IgM and IgG antibodies in sera of patients that had CMV DNA detected in CSF samples were done with use of Abbott Architect CMV IgM/IgG assays (Abbott diagnostic division), and was performed according to the manufacturer's instructions.
A chemiluminescent microparticle immunoassay (CMIA, Abbott diagnostic division, Lake Forest, Illinois 60045, USA) was used for the detection of IgM antibodies to rubella virus in serum, and was performed according to the manufacturer's instructions.
For detection of EBV antibodies in serum, an IgG/IgM ELISA (Enzygnost, Dade-Behring, Marburg, Germany) using recombinant viral capsid antigen (VCA) and EBV nuclear antigen (EBNA) was used, and was performed according to the manufacturer's instructions.
Detection of CMV-specific IgM and IgG antibodies in sera of patients that had CMV DNA detected in CSF samples were done with use of Abbott Architect CMV IgM/IgG assays (Abbott diagnostic division), and was performed according to the manufacturer's instructions.
7
Immunogenicity of Measles, Mumps, and Rubella Vaccine
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We assessed the antibody GMCs against each virus at D42 as the primary endpoint of the study. The secondary immunogenicity endpoints were: 1) the SRR at D42 (SRR was defined as an IgG antibody concentration ≥200 mIU/mL for anti-measles, ≥10 EU/mL for anti-mumps, and ≥10 IU/mL for anti-rubella irrespective of the baseline antibody concentrations; these thresholds were accepted by the USA Food and Drug Administration as offering clinical benefit); and 2) the percentage of participants with a ≥4-fold increase between D0 and D42 in GMCs for each antigen.
We determined the concentration of IgG antibodies in the blood samples taken at D0 and D42. We used commercial ELISA kits for anti-measles, anti-rubella (Enzygnost, Dade Behring), and anti-mumps antibodies (Pharmaceutical Product Development, Inc).
We determined the concentration of IgG antibodies in the blood samples taken at D0 and D42. We used commercial ELISA kits for anti-measles, anti-rubella (Enzygnost, Dade Behring), and anti-mumps antibodies (Pharmaceutical Product Development, Inc).
8
Comprehensive Hemodynamic Monitoring in Emergency Care
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Standard hemodynamic parameters were measured by both invasive and non-invasive monitoring for 360 minutes. Heart rate, respiratory rate, ETCO2, FiO2, SaO2, arterial blood pressures, pulmonary blood pressures, central venous pressure, cardiac output (CO), and ICP were recorded every 5 minutes until hospital arrival (T60) and every 15 minutes, thereafter. Pulmonary artery wedge pressure (PAWP) and urine output (UO) were measured every 15 minutes. CO was measured continuously and cerebral perfusion pressure (CPP) calculated by subtracting ICP from mean arterial pressure (MAP). Blood gas content (arterial, mixed venous and sagittal sinus) was analyzed by using an automatic analyzer (ABL 705, Radiometer, Copenhagen, Denmark) every 15 minutes until T60, then every 60 minutes, thereafter. Blood samples were also taken at T0, T15, T60, and T360 for in vitro assessments. Complete CBC (Pentra 60C+ cell counter, HORIBA ABX Diagnostics, Irvine, CA), thromboelastography (TEG) (TEG 5000 Hemostasis Analyzer, Haemoscope Corp., Niles, IL), coagulation parameters (Stat Compact, Diagnostica Stago, Parsippany, NJ), and thrombin-antithrombin (TAT) (Enzygnost, Dade Behring, Stuart, FL) were determined. The level of rFVIIa in blood samples was determined using a factor VIIa kit (Sta-FVIIa, Diagnostica Stago).
9
Anti-VZV Antibody Seroresponse Assessment
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Blood samples were collected from sub-cohorts of participants (± first 400 subjects, after randomization to the treatment groups) pre-vaccination and at 42 days post-dose 1 vaccination and 42 days post-dose 2 vaccination. Anti-VZV immunoglobulin G concentrations were measured using a commercial enzyme linked immunosorbent assay (Enzygnost, Dade Behring, Marburg, Germany) with a cut-off of 25 m-International Units [mIU]/mL. Seroresponse was defined as post-vaccination anti-VZV antibody concentration ≥ 50 mIU/mL for participants who were seronegative (antibody concentration < 25 mIU/mL) before vaccination. Anti-VZV antibody levels ≥50 mIU/mL were considered to offer clinical benefit.
10
Fibrinogen Concentrate Effects on Coagulation
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Blood was collected for analysis before injury (baseline: BL), at the end of HD, immediately before starting infusion of fibrinogen concentrate/saline (t = 0) and at 20, 40, 100, 220, 340, 700, and 1420 minutes after the start of infusion. Partial pressure of oxygen, pH, and carbon dioxide were measured with a blood gas analyzer (ABL725, Radiometer GmbH, Willich, Germany). A standard hematology analyzer (MEK-6108, Nihon Kohden, Tokyo, Japan) was used to measure platelet count. Prothrombin time (PT; reagent: Innovin), activated partial thromboplastin time (aPTT; reagent: Actin FS), and d -dimers (Innovance) were measured using the appropriate reagents and tests from Dade Behring (Marburg, Germany) on an MC 4 plus steel-ball coagulometer (Merlin Medical, Lemgo, Germany). Enzyme-linked immunosorbent assays (ELISAs), specific to porcine fibrinogen (Pig Fibrinogen; ICL Inc, Portland, Oregon) and to human fibrinogen (Human Fibrinogen; Abnova, Heidelberg, Germany), were performed. The levels of TAT were quantified by ELISA (Enzygnost, Dade Behring).
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