Method and kit for the detection of heparin-dependent...

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G01N 33/53 (2006.01) C12Q 1/56 (2006.01) G01N 33/543 (2006.01)

Patent

CA 2655582

The invention relates to a method for the detection of heparin-dependent antibodies and the diagnosis of immune or autoimmune pathologies potentiated by a heparin substance, such as thrombocytopenia induced by heparin (HIT type II) as inducer drug. The invention is characterized in that said method comprises the steps consisting in: 1) reacting: at least one substance with high affinity for a heparin substance (SFA), consisting of any molecule or complex having a high affinity for a heparin substance, with at least one heparin substance (SH) so as to form a substance with high affinity for heparin-heparin substance (SFA-SH) complex, wherein the heparin substance is in slight or large excess, so as to be able to bind other substances with affinity for heparin; then with at least one potential antigenic substance capable of reacting with the heparin substance, thus forming a complex (Ag-SH); and, finally, with the plasma or the serum of a patient to be tested (preferably diluted to a dilution of the order of 1:100) potentially containing: an anti(Ag-SH) antibody material generated in the organism after administration of a heparin substance, and/or at least one immune complex consisting of an Ag-SH/anti(Ag-SH) antibody complex present in the plasma or serum of the patient capable of reacting with the substance with high affinity for the heparin substance-heparin substance (SFA-SH) complex, and then, 2) revealing the resulting (Ag-SH)-anti(Ag-SH) complex produced.

L'invention concerne un procédé pour la détection des anticorps héparine-dépendants et le diagnostic des pathologies immunes ou auto-immunes potentialisées par une substance héparinique telle que la thrombophénie induite par l'héparine (TIH de type II) en tant que médicament inducteur. L'invention consiste en ce que ledit procédé comprend les étapes consistant à : 1) à faire réagir : au moins une substance à forte affinité pour une substance héparinique (SFA) constituée par toute molécule ou complexe ayant une forte affinité pour une substance héparinique, avec au moins une substance héparinique (SH) pour former un complexe substance à forte affinité pour l'héparine-substance héparinique (SFA-SH), la substance héparinique étant en léger ou large excès, afin de pouvoir fixer d'autres substances affines pour l'héparine; puis, avec au moins une substance antigénique potentielle capable de réagir avec la substance héparinique, formant ainsi un complexe (Ag-SH); et enfin avec le plasma ou le sérum d'un patient à tester (dilué, de préférence à une dilution de l'ordre de 1 :100) contenant potentiellement : un matériau anticorps anti(Ag-SH) généré dans l'organisme après administration d'une substance héparinique, et/ou au moins un complexe immun constitué d'un complexe Ag-SH anticorps anti(Ag-SH) présent dans le plasma ou sérum du patient capables de réagir avec le complexe substance à forte affinité pour la substance héparinique substance héparinique (SFA-SH), puis, 2) révéler le complexe produit (Ag-SH)-anti(Ag-SH) résultant.

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