the-nijmegen-hemostasis-assay-simultaneous-fluorogenic-measurement

The Nijmegen Hemostasis Assay: Simultaneous Fluorogenic Measurement of Thrombin and Plasmin Generation in a Single Well

Thrombin and plasmin both play a central role in hemostasis, regulating coagulation and fibrinolysis, respectively. Conventional hemostatic assays often ...

Hemophilia Treatment Center Nijmegen-Eindhoven-Maastricht

Radboud university medical center

Membrane-associated ATP binding cassette (ABC) transport proteins hydrolyze ATP in order to translocate a broad spectrum of substrates, from single ions to macromolecules across membranes. In humans, members from this transport family have been linked to drug resistance phenotypes, e.g., tumour resistance by enhanced export of chemotherapeutic agents from cancer cells due to gene amplifications or polymorphisms in multidrug resistance (MDR) protein 1. Similar mechanisms have linked the Plasmodium falciparum PfMDR1 transporter to anti-malarial drug resistance acquisition. In this study, the possible involvement of two related MDR proteins, PfMDR2 and PfMDR5, to emerging drug resistance is investigated by a reverse genetics approach.

PfMDR2 and PfMDR5 are dispensable for Plasmodium falciparum asexual parasite multiplication but change in vitro susceptibility to anti-malarial drugs.

Patients with moderate hemophilia express varying bleeding phenotypes.

Patients with moderate hemophilia A and B with a severe bleeding phenotype have an increased burden of disease.

Anti-factor VIII (FVIII) antibodies have been reported to exhibit both neutralizing and non-neutralizing characteristics. This is the first study investigating the full spectrum of FVIII-specific antibodies, including non-neutralizing antibodies, very-low titer inhibitors, and inhibitors, in a large nationwide population of persons with hemophilia A of all severities.

The spectrum of neutralizing and non-neutralizing anti-FVIII antibodies in a nationwide cohort of 788 persons with hemophilia A.

Thrombin generation assays (TGAs) assess the overall functionality of the hemostatic system and thereby provide a reflection of the hemostatic capacity of patients with disorders in this system. Currently, four (semi-)automated TGA platforms are available: the Calibrated Automated Thrombogram, Nijmegen Hemostasis Assay, ST Genesia and Ceveron s100. In this study, we compared their performance for detecting patients with congenital single coagulation factor deficiencies.

Comparative analysis of thrombin generation platforms for patients with coagulation factor deficiencies: A comprehensive assessment.

α2-Antiplasmin (A2AP) deficiency is a rare and often unidentified disorder characterized by increased fibrinolysis and subsequent bleeding. Global hemostasis assays may increase insight into the altered coagulation and fibrinolysis in these patients.

Enhanced thrombin and plasmin generation profiles in alpha-2-antiplasmin-deficient patients: Data from the Rare Bleeding disorders in the Netherlands study.

Congenital fibrinogen disorders (CFDs), encompassing quantitative (hypo-/afibrinogenemia) and qualitative (dysfibrinogenemia) defects, can result in bleeding or thrombotic events. This study aimed to enhance understanding of the clinical and genetic characteristics of CFD patients.

Phenotype, genotype, and laboratory assessment of congenital fibrinogen disorders: Data from the Rare Bleeding disorders in the Netherlands study.

Rare coagulation factor deficiencies (RCFDs) can cause significant bleeding, but activated partial thromboplastin time (APTT) and prothrombin time (PT) may not detect abnormalities, and factor activity measurements do not predict bleeding severity. Thrombin generation assays may better reflect overall hemostatic capacity.

Thrombin generation profiling in rare coagulation factor deficiencies: associations with bleeding severity and potential for screening.

Direct oral anticoagulants (DOACs) interfere with clot-based assays, including factor (F)VIII testing and the Nijmegen-Bethesda assay, potentially leading to false-positive results for FVIII inhibitors. Misinterpretation of these results carries serious clinical consequences. Activated charcoal-based products, such as DOAC Remove, may restore assay accuracy, but data supporting their use in FVIII inhibitor assays are limited.

Article	Year
PfMDR2 and PfMDR5 are dispensable for Plasmodium falciparum asexual parasite multiplication but change in vitro susceptibility to anti-malarial drugs. Malaria journal\| PubMed ID: 25884516	2015
Patients with moderate hemophilia A and B with a severe bleeding phenotype have an increased burden of disease. Journal of thrombosis and haemostasis : JTH\| PubMed ID: 37813197	2024
The spectrum of neutralizing and non-neutralizing anti-FVIII antibodies in a nationwide cohort of 788 persons with hemophilia A. Frontiers in immunology\| PubMed ID: 38455035	2024
Comparative analysis of thrombin generation platforms for patients with coagulation factor deficiencies: A comprehensive assessment. Thrombosis research\| PubMed ID: 38834002	2024
Enhanced thrombin and plasmin generation profiles in alpha-2-antiplasmin-deficient patients: Data from the Rare Bleeding disorders in the Netherlands study. Research and practice in thrombosis and haemostasis\| PubMed ID: 39628652	2024
Phenotype, genotype, and laboratory assessment of congenital fibrinogen disorders: Data from the Rare Bleeding disorders in the Netherlands study. Thrombosis research\| PubMed ID: 40203550	2025
Thrombin generation profiling in rare coagulation factor deficiencies: associations with bleeding severity and potential for screening. Journal of thrombosis and haemostasis : JTH\| PubMed ID: 41443372	2025
Direct oral anticoagulant interference and removal in the factor VIII inhibitor assay. Research and practice in thrombosis and haemostasis\| PubMed ID: 41624237	2026
Monitoring the Hemostatic Balance: measuring thrombin generation in a patient with acquired hemophilia A on combined pro- and anticoagulant therapy. Thrombosis research\| PubMed ID: 41650515	2026

Sanna R. Rijpma

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