A multi-center evaluation of TECHNOSCREEN ADAMTS-13 activity assay as a screening tool for detecting deficiency of ADAMTS-13.

Background: Quantifying A disintegrin-like and metalloprotease with thrombospondin type 1 motif, member 13 (ADAMTS-13) activity enhances thrombotic thrombocytopenic purpura (TTP) diagnosis but most assays are time consuming, technically demanding, and mainly available in reference centers.

Objective: Evaluate a simple, semiquantitative ADAMTS-13 activity screening test for early identification/exclusion of TTP.

Patients/methods: Plasma from 220 patients with suspected thrombotic microangiopathy at three reference centers were tested with TECHNOSCREEN ADAMTS13 activity screening test in comparison with TECHNOZYM ADAMTS-13 activity ELISA at two centers, and in-house fluorescence resonance energy transfer assay at the third center.

Development of a novel, rapid assay for detection of heparin-binding defect antithrombin deficiencies: the heparin-antithrombin binding (HAB) ratio.

Introduction: Qualitative and quantitative antithrombin deficiency predisposes to thrombosis, although patients with heparin-binding dysfunction have a lower incidence than other sub-types. Assays discriminating between qualitative sub-types are not widely available.

Patients/methods: Extended heparin incubation in antithrombin activity assays can overestimate levels in patients with HBDs.

Cellular and molecular basis of von Willebrand disease: studies on blood outgrowth endothelial cells.

Von Willebrand disease (VWD) is a heterogeneous bleeding disorder caused by decrease or dysfunction of von Willebrand factor (VWF). A wide range of mutations in the VWF gene have been characterized; however, their cellular consequences are still poorly understood. Here we have used a recently developed approach to study the molecular and cellular basis of VWD.

Denaturing HPLC for mutation screening.

Denaturing High-Performance Liquid Chromatography (dHPLC) is probably the most versatile and one of the most widely used mutation screening technologies. It benefits from a combination of relative technical simplicity and a very high sensitivity (mutation detection rate), approaching 100%. DHPLC can reliably detect single-base mismatches in fragments between 150 and 500 bp, although detection in fragments up to 1,500 bp has been reported.

The significance of published polymorphisms in 14 cases of mild factor VII deficiency.

Factor VII (FVII) plays a critical role in the initiation of blood coagulation, and patients with dysfunctional or reduced levels of this protein are susceptible to mucosal bleeding. There is poor correlation between the clinical presentation and the phenotypic data; and in cases of a mild bleeding tendency, mild to moderate reductions in both FVII antigen and activity may be overlooked. The prevalence of FVII deficiency may therefore be underestimated.

Germline mosaicism resulting in the transmission of severe hemophilia B from a grandfather with a mild deficiency.

We report a family in which the normal pattern of X-linked inheritance of hemophilia B (Factor IX deficiency) is complicated by mosaicism in the proband's maternal grandfather. The proband, an infant with severe Factor IX deficiency, was initially thought to be a sporadic case. Testing of other family members identified his mother as a carrier of the disorder, and his asymptomatic maternal grandfather as having very mild FIX deficiency.

Eighteen unrelated patients with factor XI deficiency, four novel mutations and a 100% detection rate by denaturing high-performance liquid chromatography.

Factor XI (FXI) deficiency is an autosomal bleeding disorder of variable severity. Inheritance is not completely recessive as heterozygotes may display a distinct, if mild, bleeding tendency. Eighteen unrelated FXI-deficient patients were screened blind by fluorescent single-stranded conformation polymorphism (F-SSCP) analysis and denaturing high-performance liquid chromatography (dHPLC).