Evaluation of a modified APTT-based method for determination of APC resistance in plasma from patients on heparin or oral anticoagulant therapy.

An APTT-based kit method (Coatest APC Resistance), modified by predilution 1+4 of sample plasma in a plasma diluent containing a heparin antagonist (V-DEF plasma), has been evaluated on plasmas from patients treated with unfractionated (n = 110) or either of three different low molecular heparins (n=44), or with oral anticoagulants (n=147). Irrespective of treatment, no difference was observed in the APC response as compared to untreated individuals (n=62), and a complete discrimination was obtained between individuals with a normal factor V genotype and those carrying the FV:Q506 mutation. Furthermore, in contrast to the original, APTT-based kit method, where anticoagulant therapy results in a prolongation of the APTT, the modified kit provided APTT values within the normal range for orally anticoagulated (INR< or =6) and for all heparin treated (< or =1 IU/mL) patients except for one with a suspected presence of phospholipid antibodies. Due to the predilution in V-DEF plasma, contamination with platelets up to 1.5 x 10(4)/microL had a negligible effect on analysis of frozen plasmas regarding their classification as normal or abnormal. Analyses of fresh plasmas show no influence at platelet counts up to 6x10(4)/microL. Consequently, negligible differences in APC ratios were obtained between fresh and frozen plasmas. In conclusion, the modified kit method is applicable to plasmas from anticoagulated patients as well as from untreated individuals, allowing a safe assignment regarding the presence or absence of the FV:Q506 genotype.