Paul-Brent Peta-Anne, Cattley Trudi N, Myers Stephen P, Brooks Lyndon, Cheras Phillip A
Australian Centre for Complementary Medicine Education and Research, University of Queensland and Southern Cross University, Brisbane, Queensland, Australia.
Blood Coagul Fibrinolysis. 2004 Apr;15(3):273-8. doi: 10.1097/00001721-200404000-00014.
A novel and precise assay that facilitates high-throughput screening of fibrinolytic agents was developed based on the automated assessment of the euglobulin clot lysis time in microtitre plates. Euglobulin fractions from fresh plasma samples were assessed over 28 days to determine the inter-assay and intra-assay precision. The intra-assay (coefficient of variation range, 0.7-2.6%) and inter-assay precision (coefficient of variation range, 6.8-12.1%) was found to be well within limits required by the Food and Drug Administration. On day 1 and day 28, the results of the microtitre plate euglobulin clot lysis time method were compared with tissue plasminogen activator activity, plasminogen activator inhibitor activity and results produced on fibrin plates. All comparisons were found to correlate significantly. The validity of this method for assaying fibrinolytic agents was assessed by comparing dose-response curves for streptokinase produced using fibrin plates and this method. The critical influence of ambient temperature on the inter-assay reproducibility of this method was established by testing samples over a range of temperatures between 20 degrees C and 40 degrees C.
基于微量滴定板中优球蛋白凝块溶解时间的自动评估，开发了一种新颖且精确的检测方法，便于对纤溶药物进行高通量筛选。对新鲜血浆样本中的优球蛋白组分进行了28天的评估，以确定批间和批内精密度。发现批内精密度（变异系数范围为0.7 - 2.6%）和批间精密度（变异系数范围为6.8 - 12.1%）完全在食品药品监督管理局要求的限度内。在第1天和第28天，将微量滴定板优球蛋白凝块溶解时间法的结果与组织纤溶酶原激活物活性、纤溶酶原激活物抑制剂活性以及纤维蛋白平板上产生的结果进行了比较。发现所有比较均具有显著相关性。通过比较使用纤维蛋白平板和该方法产生的链激酶剂量 - 反应曲线，评估了该方法检测纤溶药物的有效性。通过在20℃至40℃的一系列温度下测试样本，确定了环境温度对该方法批间重现性的关键影响。
