凝固时间法测定血浆AT-Ⅲ活性

Estabilishment of the Clotting-Time Method for Determination of Antithrombin III Activity in Human Plasma

目的介绍凝固时间法测定血浆AT-Ⅲ活性的实验方法。方法将校正血浆用含肝素缓冲液稀释成不同浓度(即不同活性AT-Ⅲ),加入纯品纤维蛋白原共同孵育后,测定其凝固时间;根据不同AT-Ⅲ活性与相应凝固时间制作标准曲线;测定待检血浆凝固时间,查标准曲线即可得到待检血浆AT-Ⅲ活性。结果该法AT-Ⅲ活性在25%～125%范围内线性良好;日内变异系数为3.2%,日间变异系数为4.9%;凝固时间法与发色底物法比较相关性良好(r=0.999,P<0.05);检测178例健康人血浆AT-Ⅲ活性为105.4%±18.8%。结论该方法重复性好,结果可靠,简便实用,可用于常规检验。

Objective: To establish a method for determination of antithrombin III (AT-III) activity in human plasma.Method: The plasma with a series activity of AT-III was incubated with pure fibrinogen,then the clotting time were measured by coagulovisimeter.The standard curve was established between AT-III activity and respective clotting time.To calculate the AT-III activity of detecting plasma by compared with clotting thime in the standard curve.Results: A good linear relationship was obtained between AT-III activity and clotting time in the range of 25%～150% activity. The intra-day CV was 3.2% and between-run was 4.9%. A good correlation was found when compared with chromogenic substrate method (r=0.999,P<0.05). The AT-III activity was 105.4%±18.8% in 178 healthy subjects.Conclusion: It is simple, repeatable, reliable, practical and suitable for routine clinical test.