Venous blood was collected in EDTA tubes. The whole-plasma samples were stored at −80°C until analyses were performed. Laboratory analyses (lipid, glucose, creatinine, CD4 and CD8 cells, HIV copies and RNA levels) were performed according to routine practice in our hospital. sTWEAK and sCD163 plasma concentrations were measured in duplicate with commercially available ELISA kits (Bender MedSystems, [Vienna, Austria]; and R&D Systems [Abingdon, UK], respectively). hsCRP, sTNFRII, sVCAM-1 and ADMA levels were also determined with commercially available ELISA kits (Immundiagnostik AG [Bensheim, Germany], R&D Systems [Abingdon, UK], DLD Diagnostika GmbH [Hamburg, Germany] and eBioscience's [San Diego, USA], respectively). IL-6 was measured by ultrasensitive ELISA (Quantikine HS Human IL-6 Immunoassay; R&D Systems [Abingdon, UK]). D dimer levels were measured in an automated latex-enhanced immunoassay (HemosIL D-Dimer HS 500; Instrumentation Laboratory [Bedford, UK]). Ig G antibodies to cytomegalovirus were determined by enzyme immunoassay (GenWay Biotech [San Diego, USA]).
Hemosil d dimer hs 500
Manufactured by Werfen
Sourced in United Kingdom, Switzerland, Germany, Spain, India
The HemosIL D-Dimer HS 500 is a laboratory equipment product designed for the quantitative determination of D-Dimer in human plasma. It is a diagnostic tool used to assist in the assessment of thrombotic disorders.
Automatically generated - may contain errors
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14 protocols using hemosil d dimer hs 500
1
Biomarker Profiling in Blood Samples
2
Immunological and Viral Biomarker Measurements
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CD4 T-cell counts were determined in fresh whole blood using an Epic XL-MCL flow cytometer (Beckman-Coulter, Brea, CA, USA) according to the manufacturer’s instructions. Plasma HIV-1 RNA concentration was measured using quantitative polymerase chain reaction (COBAS Ampliprep/COBAS Taqman HIV-1 test, Roche Molecular Systems, Basel, Switzerland) according to the manufacturer’s protocol. The detection limit for this assay was 20 HIV RNA copies per milliliter.
High-sensitive C-reactive protein (hsCRP) and β2-microglobulin were determined with an immunoturbidimetric assay using COBAS 701 (Roche Diagnostics, GmbH, Mannheim, Germany).d -dimer levels were determined using an automated latex enhanced immunoassay (HemosIL, d -Dimer HS 500, Instrumentation Laboratory) in plasma samples stored at −20°C.
High-sensitive C-reactive protein (hsCRP) and β2-microglobulin were determined with an immunoturbidimetric assay using COBAS 701 (Roche Diagnostics, GmbH, Mannheim, Germany).
3
COVID-19 D-Dimer Levels and Outcomes
4
D-Dimer Quantification in CSF and Blood
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The quantitative determination of D-dimer in citrated CSF was performed using a Luminescent Oxygen Channeling Immunoassay (INNOVANCE LOCI) based on an Atellica COAG 360 System (Siemens Healthineers, Erlangen, Germany). The calibration curve showed a linear relationship between the plotting signal and the analyte concentration (hs D-Dimer INNOVANCE LOCI) with only minimal deviation from the manufacturer's target value (max. 9.4%). Dilution series were performed to assess measurability of D-dimer in very low concentrations (1:10 [22 ng/mL], 1:20 [11 ng/mL], 1:50 [4 ng/mL], 1:100 [2 ng/mL], and 1:200 [1 ng/mL]). Intra-assay imprecision of the assay was between 0.66% and 0.73 %CV using 3 individual samples (high control, low control, and plasma pool) in 20 replicates. Duplicate testing in 24 randomly selected patients confirmed the accuracy of the test results. All laboratory tests were performed in a single run within 1 day and under identical circumstances. The quantitative determination of D-dimer in citrated blood plasma was performed with a HemosIL D-Dimer HS 500 latex-enhanced immunoassay on an ACL TOP 700 testing system (Instrumentation Laboratory, Bedford, MA).
5
COVID-19 D-Dimer Surveillance in Hospitalized Patients
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Data were obtained from the electronic health record (Epic Systems, Verona, WI), which is an integrated electronic health record including all inpatient and outpatient visits in the health system.
Patients hospitalized with a positive polymerase chain reaction test for COVID-19 were eligible for this retrospective, observational study if ≥1 D-dimer was measured during hospital admission. At all 4 NYU Langone Health inpatient facilities, routine D-dimer surveillance for individuals with suspected or confirmed diagnoses of COVID-19 was included in COVID-19-specific admission order sets in the electronic heath record at the time of hospital admission starting March 25. At all NYU Langone Health sites, D-dimer assay was measured using the Hemosil D-dimer HS 500 on an automated coagulation analyzer (ACL TOP, Instrumentation Laboratory). The initial D-dimer and all D-dimers measured during hospital admission were recorded for all eligible patients. The upper limit of normal for the D-dimer assay is 230 ng/mL. Subjects were categorized into normal (D-dimer <230 ng/mL) and elevated (D-dimer ≥230 ng/mL) categories. We conducted sensitivity analyses using different D-Dimer categories: <230 ng/mL (normal), 230 to 500 ng/mL, 500 to 2000 ng/mL, and >2000 ng/mL.
Patients hospitalized with a positive polymerase chain reaction test for COVID-19 were eligible for this retrospective, observational study if ≥1 D-dimer was measured during hospital admission. At all 4 NYU Langone Health inpatient facilities, routine D-dimer surveillance for individuals with suspected or confirmed diagnoses of COVID-19 was included in COVID-19-specific admission order sets in the electronic heath record at the time of hospital admission starting March 25. At all NYU Langone Health sites, D-dimer assay was measured using the Hemosil D-dimer HS 500 on an automated coagulation analyzer (ACL TOP, Instrumentation Laboratory). The initial D-dimer and all D-dimers measured during hospital admission were recorded for all eligible patients. The upper limit of normal for the D-dimer assay is 230 ng/mL. Subjects were categorized into normal (D-dimer <230 ng/mL) and elevated (D-dimer ≥230 ng/mL) categories. We conducted sensitivity analyses using different D-Dimer categories: <230 ng/mL (normal), 230 to 500 ng/mL, 500 to 2000 ng/mL, and >2000 ng/mL.
6
D-dimer Assay Cutoffs for VTE Risk
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D-dimer concentrations were assessed with the quantitative assay routinely used in each participating center, provided it was one of the following: Cobas h232 (Roche Diagnostics; Switzerland), HemosIL D-dimer HS 500 (Instrumentation Laboratory; Milan, Italy), HemosIL D-dimer HS (Instrumentation Laboratory), HemosIL D-dimer (Instrumentation Laboratory), Innovance D-DIMER (Siemens; Deerfield, IL), Sclavo Auto D-dimer (Dasit; Milan, Italy), STA Liatest D-dimer (Diagnostica Stago; Asnieres-sur-Seine, France), and VIDAS D-dimer Exclusion (bioMerieux; Lyon, France). For the assays expressing results as fibrinogen equivalent units, the cutoffs were 350 ng/mL and 500 ng/mL for males and females, respectively; the cutoffs for assays expressing the results as D-dimer units were 175 ng/mL and 250 ng/mL for males and females, respectively. These cutoffs were selected by the study team after a critical evaluation of the Dulcis study results,7 (link) taking into account that (1) patients aged ≥75 years were excluded from the present study (and therefore, there was no need for different cutoff levels according to the age) and (2) male patients are unanimously recognized at higher risk of recurrence than females (for this reason the decided cutoff level was lower for males than females).
7
Hematological and Biochemical Profiles in Patients
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Hematological and biochemical profiles were evaluated in all patients upon admission to each hospital. Among them, CRP was determined by an immunoturbidimetric method (Cobas 701; Roche Diagnostics, Mannheim, Germany), D-dimer by an automated latex-enhanced immunoassay (HemosIL D-Dimer HS 500, Instrumentation Laboratory, Bedford, Massachusetts), LDH and ferritin were determined by enzyme-linked immunosorbent assay (ELISA).
8
Biochemical Marker Assessment Protocol
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Biochemical soluble markers, including blood proteins, micronutrients, soluble iron metabolism-related markers, and inflammatory biomarkers, were routinely determined by standard procedures at the Biochemistry Service of our hospital. Briefly, total iron, transferrin, and soluble transferrin receptor (sTfR) were measured by photometry and ferritin by particle enhanced immunoturbidimetric assay in a Hitachi Cobas C702 modular analyzer (Roche Diagnostics, Rotkreuz, Switzerland). hsCRP and β2-microglobulin levels were determined in frozen serum samples with an immunoturbidimetric assay using Cobas 701 (Roche Diagnostics, Mannheim, Germany). Measurements of the levels of homocysteine were performed by photometry according to the manufacturer’s instructions, and D-dimer levels were quantified by using an automated latex enhanced immunoassay using frozen plasma samples (HemosIL D-Dimer HS 500, Instrumentation Laboratory, Bedford, Massachusetts).
9
Plasma D-dimer Measurement for VTE Exclusion
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Blood samples were collected in 3.8% (0.129 M) sodium citrate (anticoagulant) tubes (BD Vacutainer) and plasma was obtained by 10-minute centrifugation at 3000 rpm. All plasma samples were analyzed within two hours of collection. Plasma Ddimer concentration was measured using a latex-enhanced immunoassay (Hemosil D-Dimer HS 500, Instrumentation Laboratory) on an automated coagulation analyzer (ACL TOP 750, Instrumentation Laboratory). D-dimer levels were expressed in fibrinogen equivalent units (FEUs). At our institution, the cutoff established for VTE exclusion was 0.5 μg/mL (FEU).
10
Standardized Coagulation Assays and Antibody Testing
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Standardized coagulation assays in citrated (3.2%)-plasma were as follows: prothrombin time (PT, Medirox Owren's PT, Medirox, Nyköping, Sweden), fibrinogen (Clauss method, HemosIL Q.F.A. Thrombin, Werfen, Barcelona, Spain), and D-dimer (HemosIL D-Dimer HS 500). Antibodies (classes IgG, IgM and IgA) against PF4-heparin complexes were tested both with a gel agglutination rapid assay (ID-PaGIA Heparin/PF4 Antibody Test, Bio-Rad Laboratories, USA) and an enzyme-linked immunosorbent assay (ELISA) (Asserachrom HPIA, Diagnostica Stago, France). ELISA (Asserachrom HPIA, Diagnostica Stago) results were reported as percentage of absorbance related to reference sample. The reference sample was within the manufacturer specifications (stated on package insert, dependent on reagent lot). Percentage result is calculated (sample absorbance/reference absorbance). The interpretation of positivity is as follows: < 30 % of reference; negative, 30-50%, borderline positive, >50% positive, and >100% strongly positive.These results are referred below as anti-PF4 antibodies [1] (link). Cardiolipin and beta-2-glycoprotein-I IgG antibodies were analyzed using Phadia 250 immuno-analyzer and reagents (Sweden).
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