Limit of detection of latex agglutination test was determined by testing VWF antibody-latex reagent on low-level samples (cryosupernatant). Seven cryosupernatant samples were serially diluted twofold, and the VWF : Ag levels in the samples (neat) were measured by immunoturbidimetric assay using Hemosil von Willebrand Factor Antigen kit (Instrumentation Laboratory, Italy). VWF : Ag levels in 1 : 2, 1 : 4, and 1 : 8 diluted samples were estimated by dividing the VWF : Ag levels of neat samples by the dilution factors of 2, 4, and 8, respectively. Latex agglutination test was performed to determine the lowest level of VWF : Ag that shows positive agglutination reaction.
Hemosil von willebrand factor antigen kit
Manufactured by Werfen
Sourced in Italy, United States
The Hemosil von Willebrand Factor Antigen kit is a laboratory diagnostic tool used to measure the levels of von Willebrand factor antigen in patient samples. It is designed to aid in the diagnosis and monitoring of von Willebrand disease and other related coagulation disorders.
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5 protocols using hemosil von willebrand factor antigen kit
1
Determining Limit of Detection for Latex Agglutination Test
2
Factor VIII Activity and Inhibitor Assay
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FVIII activity was detected using a one-stage activated partial thromboplastin time (aPTT)-based assay (three dilutions of each sample), against an international plasma standard. All measurements were performed on an ACL TOP-500 analyzer (Instrumentation Laboratory, Bedford, MA, USA). The reagents of FVIII:C assay included HemosIL SynthASil and Calcium Chloride, HemosIL FVIII Deficient Plasma, HemosIL Calibration Plasma, HemosIL Normal Control 1 Unassayed, HemosIL Special Test Control Level 2, HemosIL Factor Diluent (Instrumentation Laboratory, Bedford, MA, USA). VWF:Ag determinations were also performed on an ACL TOP-500 analyzer with a latex particle-enhanced immunoturbidimetric assay using HemosIL von Willebrand factor antigen kit (Instrumentation Laboratory, Bedford, MA, USA). The Nijmegen modification of the Bethesda assay was used for determination of FVIII inhibitors.
3
Linearity of Latex Agglutination Test
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The linearity of latex agglutination test was determined by testing VWF antibody-latex reagent on two patient samples which were serially diluted twofold. The VWF : Ag level in diluted samples was measured by immunoturbidimetric assay using Hemosil von Willebrand Factor Antigen kit (Instrumentation Laboratory, Italy). Semiquantitative latex agglutination test was performed on diluted samples, and the end-titres were compared with the VWF : Ag levels. Graphical presentations of the semiquantitative VWF : Ag by latex agglutination test (Y) versus the VWF : Ag by immunoturbidimetric assay (X) were made for individual sample and also for a combination of the two samples.
4
Factor VIII Activity and Inhibitor Measurement
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The one‐stage‐based activated partial thromboplastin time assay (Instrumentation Laboratory, Bedford, MA, USA) was used to measure FVIII activity in a multi‐dilution mode, with FVIII‐deficient plasma. The Nijmegen modification of the Bethesda Assay was used for the determination of FVIII inhibitors. An ACL TOP‐700 analyzer was used to perform all measurements (Instrumentation Laboratory, Bedford, MA, USA). A latex particle‐enhanced immunoturbidimetric assay with HemosIL von Willebrand factor antigen kit (Instrumentation Laboratory, Bedford, MA, USA) was utilized to determine VWF:Ag levels.
5
Comparative Analysis of VWF:Ag Measurement
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Method comparison study between latex agglutination test and the reference method (immunoturbidimetric assay) was conducted by parallel analysis of 132 plasma samples of different levels of VWF : Ag. The samples were obtained from 40 healthy individuals, 40 patients with cardiovascular diseases, and 32 patients with other diseases including malignant and inflammatory disorders. The remaining 20 samples were cryosupernatant samples (i.e., samples with low levels of VWF : Ag) that were obtained from Blood Transfusion Services. All specimens were tested in ten days, and the testing was spread over ten months.
Qualitative and semiquantitative latex agglutination tests were performed using the VWF antibody-latex reagents. The immunoturbidimetric assay was performed on ACL Elite Pro Coagulation analyser (Instrumentation Laboratory, Italy) using Hemosil von Willebrand Factor Antigen kit from Instrumentation Laboratory, Italy. The procedure for VWF : Ag by immunoturbidimetric assay was performed according to the manufacturer's instructions and adhered to the CLSI Document H21-A5 on the specimen collection and processing of blood specimens for testing the plasma-based coagulation assays. Normal reference range for VWF : Ag levels by immunoturbidimetric assay in our laboratory is 50–150%.
Qualitative and semiquantitative latex agglutination tests were performed using the VWF antibody-latex reagents. The immunoturbidimetric assay was performed on ACL Elite Pro Coagulation analyser (Instrumentation Laboratory, Italy) using Hemosil von Willebrand Factor Antigen kit from Instrumentation Laboratory, Italy. The procedure for VWF : Ag by immunoturbidimetric assay was performed according to the manufacturer's instructions and adhered to the CLSI Document H21-A5 on the specimen collection and processing of blood specimens for testing the plasma-based coagulation assays. Normal reference range for VWF : Ag levels by immunoturbidimetric assay in our laboratory is 50–150%.
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