Rodgers Susan E, Wong Amanda, Gopal Reanuga D, Dale Brian J, Duncan Elizabeth M, McRae Simon J
Haematology Division, IMVS, SA Pathology, Adelaide, SA, Australia; School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, SA, Australia.
School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, SA, Australia.
Thromb Res. 2014 Jul;134(1):160-4. doi: 10.1016/j.thromres.2014.04.010. Epub 2014 Apr 22.
There is minimal data on the influence of pre-analytical variables on the use of calibrated automated thrombography (CAT), to measure thrombin generation.
To evaluate the impact of centrifugation methods, time after collection, and contact activation inhibition on the CAT assay performed using two commercial reagents.
Six different methods of plasma separation were examined. Thrombin generation triggered by a 5 pM tissue factor reagent was not greatly affected by plasma separation method, with similar results obtained with all methods apart from single centrifugation and membrane filtration. Membrane filtration increased APTT and is not recommended. Extended double centrifugation at higher speed was required to minimise the impact of residual phospholipid with 1 pM tissue factor trigger, particularly with inhibition of contact activation. The effect of a delay of up to 24 hours in preparing plasma was assessed. No significant difference in results was observed among samples processed between 0.5 and 6 hours after blood collection into plastic Vacuette® tubes. The presence or absence of corn trypsin inhibitor had a significant impact on all parameters with 1 pM tissue factor trigger, with minor differences seen on Peak and ttPeak results using 5 pM tissue factor.
The impact of pre-analytical variables on thrombin generation results is dependent on the concentration of tissue factor in the trigger reagent used. Results with 1 pM tissue factor are particularly sensitive to centrifugation method and contact activation, and standardisation is required to allow large collaborative studies to be performed.
关于分析前变量对校准自动血栓形成测定法（CAT）用于测量凝血酶生成的影响的数据极少。
评估离心方法、采血后时间以及接触激活抑制对使用两种商业试剂进行的CAT测定的影响。
检查了六种不同的血浆分离方法。由5 pM组织因子试剂触发的凝血酶生成受血浆分离方法的影响不大，除了单次离心和膜过滤外，所有方法得到的结果相似。膜过滤会增加活化部分凝血活酶时间（APTT），不建议使用。对于1 pM组织因子触发物，需要更高速度的延长双离心以最小化残留磷脂的影响，特别是在接触激活受到抑制时。评估了血浆制备延迟长达24小时的影响。在血液采集到塑料Vacuette®管后0.5至6小时内处理的样本之间，未观察到结果有显著差异。玉米胰蛋白酶抑制剂的存在与否对1 pM组织因子触发物的所有参数有显著影响，使用5 pM组织因子时，在峰值和ttPeak结果上有微小差异。
分析前变量对凝血酶生成结果 的影响取决于所用触发试剂中组织因子的浓度。1 pM组织因子的结果对离心方法和接触激活特别敏感，需要标准化以进行大型协作研究。
