Enzyme-linked coagulation assay. II. A sensitive assay for tissue factor and factors II, VII, and X.
Enzyme-linked coagulation assay. II. A sensitive assay for tissue factor and factors II, VII, and X.
复制标题
酶联凝血测定。
DOI：
10.1016/0003-2697(86)90141-7
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发表时间：
1986
影响因子：
2.9
通讯作者：
Rothberger,H
中科院分区：
文献类型：
--
作者：
Doellgast,GJ;Rothberger,H
我们已经开发了一种固相凝血试验，它使用过氧化物酶-纤维蛋白原溶液和纤维蛋白原结合到微量滴定板作为从凝血级联产生的凝血酶的底物。我们已经开发了这种检测方法，用于测量外源性途径因子凝血活酶（组织因子，因子III），VII和VIIa，X和II。使用长的孵育时间（40-90分钟），凝血活酶可以在非常低的浓度下在人脑提取物中测量。通过显示对因子II、V、X和VII的要求，但不对VIII、IX、XI或XII的要求，证明了促凝血酶原激酶的特异性;在证明该特异性时，使用了单一因子和纯因子混合物中单一缺陷的底物血浆。对该试验进行了修改，以使用适当的缺陷血浆测量因子II、VII、VIIa和X。检测限为2-3个数量级低于一个阶段的凝血试验测定的所有因素。该检测方法具有方便、特异性与标准凝血试验相当、灵敏度高的优点。
We have developed a solid-phase clotting assay which uses peroxidase-fibrinogen in solution and fibrinogen bound to microtiter plates as a substrate for the thrombin generated from the clotting cascade. We have developed this assay for measurement of the extrinsic pathway factors thromboplastin (tissue factor, factor III), VII and VIIa, X, and II. Using long incubation times (40–90 min), thromboplastin could be measured in extracts of human brain at very low concentrations. Specificity for thromboplastin was demonstrated by showing a requirement for factors II, V, X, and VII but not for VIII, IX, XI, or XII; both substrate plasmas monodeficient in single factors and mixtures of the pure factors were used in demonstrating this specificity. The assay was modified to measure factors II, VII, VIIa, and X using appropriate deficient plasmas. The limit of detection was 2–3 orders of magnitude lower than a one-stage clotting test for all factors assayed. This assay has the advantages of convenience, specificity comparable to standard clotting tests, and high sensitivity.