D'Alessandro Elisa, Scaf Billy, van Oerle René, van Nieuwenhoven Frans A, van Hunnik Arne, Verheule Sander, Schotten Ulrich, Ten Cate Hugo, Spronk Henri M H
Department of Biochemistry and Internal Medicine Cardiovascular Research Institute Maastricht Maastricht University Medical Center Maastricht The Netherlands.
Department of Physiology Cardiovascular Research Institute Maastricht Maastricht University Medical Center Maastricht The Netherlands.
Res Pract Thromb Haemost. 2021 Nov 21;5(8):e12620. doi: 10.1002/rth2.12620. eCollection 2021 Dec.
The goat model of atrial fibrillation (AF) allows investigation of the effect of AF on coagulation. However, assays for goat plasma are not available from commercial sources. Calibrated automated thrombography (CAT) provides a global view of the coagulation profile by assessing thrombin generation (TG). We describe the customization of the CAT assay in goat platelet-poor plasma (PPP) and in factor Xa (FXa)-inhibitor-anticoagulated PPP. TG was initiated in the presence of phospholipids and either (a) PPP reagent, reagent low, or reagent high; (b) goat brain protein extraction (GBP); or (c) Russell's viper venom-factor X activator (RVV-X). Contact activation was assessed by adding corn trypsin inhibitor. Different concentrations of prothrombin complex concentrate (PCC) were used to determine the sensitivity of both the GBP and RVV-X method. To obtain FXa-inhibitor anticoagulated plasma, rivaroxaban was added to plasma. TG settings with human reagents were not suitable for goat plasma. TG triggered with GBP increased peak height and ETP values. Similarly, the RVV-X method produced comparable TG curves and was more sensitive to PCC titration. Finally, both methods were able to detect the decrease in clotting potential induced by FXa inhibition. This is the first study that reports the customization of the CAT assay for goats. The GBP and RVV-X methods were comparable in triggering TG in goat plasma. The RVV-X method seemed to better discriminate changes in TG curves due to increases in clotting potential as well as to FXa inhibition by rivaroxaban in goat plasma.
心房颤动（AF）的山羊模型可用于研究AF对凝血的影响。然而，市面上没有可用于山羊血浆的检测方法。校准自动血栓形成测定法（CAT）通过评估凝血酶生成（TG）提供凝血概况的全局视图。我们描述了在山羊乏血小板血浆（PPP）和因子Xa（FXa）抑制剂抗凝的PPP中对CAT测定法的定制。TG在磷脂存在下由以下物质引发：（a）PPP试剂、低试剂或高试剂；（b）山羊脑蛋白提取物（GBP）；或（c）罗素蝰蛇毒因子X激活剂（RVV-X）。通过添加玉米胰蛋白酶抑制剂评估接触激活。使用不同浓度的凝血酶原复合物浓缩物（PCC）来确定GBP和RVV-X方法的敏感性。为获得FXa抑制剂抗凝血浆，将利伐沙班添加到血浆中。用人试剂的TG设置不适用于山羊血浆。用GBP触发的TG增加了峰值高度和ETP值。同样，RVV-X方法产生了可比的TG曲线，并且对PCC滴定更敏感。最后，两种方法都能够检测到由FXa抑制引起的凝血潜能降低。这是第一项报道为山羊定制CAT测定法的研究。GBP和RVV-X方法在触发山羊血浆中的TG方面具有可比性。RVV-X方法似乎能更好地区分由于凝血潜能增加以及利伐沙班对山羊血浆中FXa抑制导致的TG曲线变化。
