A three-stage chromogenic assay for the quantitative determination of unfractionated and low molecular weight heparins in human plasma by measurement of Factor Xa inhibition.The inhibitory effect of antithrombin III (ATIII) on Factor IIa (thrombin), Factor Xa (fXa) and other coagulation serine proteases in plasma is increased several-thousand fold by heparin. This inhibition accounts for the anticoagulant effect of heparin.In the first stage, heparin in the patient sample is allowed to react with ATIII. In the second stage, the heparin-ATIII complex formed reacts with excess fXa. In the final stage, the residual fXa activity hydrolyzes the fXa-specific chromogenic substrate SPECTROZYME® fXa. The amount of residual fXa activity is inversely proportional to the heparin concentration in the patient plasma.
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