Development of a rapid and sensitive chromogenic heparin assay for clinical use.
A new sensitive chromogenic heparin assay is developed, which is well suited for clinical use. For the assay two reaction mixtures are required which can be lyophilized and reconstituted on the day of use. These reagents are stable during at least 6 h. Only two time-dependent pipetting steps are necessary. Any compound that inactivates thrombin, or can potentiate thrombin inactivation by an inhibitor, can be measured with this assay, including standard heparin, low molecular weight heparins, hirudin, alpha-NAPAP, pentosan polysulphate and dermatan sulphate. It is shown that heparin can be measured accurately in whole blood and in plasma. By addition of dextran sulphate to one of the reagents a platelet factor 4-insensitive assay is developed, so heparin can be measured even in blood that is partially activated and thus contains platelet factor 4 which neutralizes heparin.
Wagenvoord
R J
RJ
Department of Biochemistry, University of Limburg, Maastricht, The Netherlands.
Hendrix
H H
HH
Kolde
H J
HJ
Hemker
H C
HC
D000925:Anticoagulants / Q000097:blood; D000990:Antithrombin III; D001779:Blood Coagulation Factors; D002122:Calcium Chloride; D002138:Calibration; D002863:Chromogenic Compounds; D003871:Dermatan Sulfate / Q000097:blood; D016264:Dextran Sulfate / Q000494:pharmacology; D004151:Dipeptides; D004355:Drug Stability; D015949:Factor IXa; D006493:Heparin / Q000097:blood*; D006801:Humans; D009842:Oligopeptides; D010743:Phospholipids; D010978:Platelet Factor 4 / Q000037:antagonists & inhibitors; D012680:Sensitivity and Specificity
D000925:Anticoagulants; D001779:Blood Coagulation Factors; D002863:Chromogenic Compounds; D004151:Dipeptides; D009842:Oligopeptides; D010743:Phospholipids; D003871:Dermatan Sulfate; D010978:Platelet Factor 4; C021129:S 2238; C031023:S 2337; C028249:N(alpha)-phosphorylalanylproline; D000990:Antithrombin III; D006493:Heparin; D016264:Dextran Sulfate; D015949:Factor IXa; D002122:Calcium Chloride