Ruzzenente O, Ghidini R, Poli G, Schiavon R, Guidi G C
Laboratory of Clinical Chemistry, C.O.C., Valeggio, Italy.
Thromb Res. 1992 Jan 15;65(2):281-7. doi: 10.1016/0049-3848(92)90248-9.
HC II functional assays are generally preferred with respect to immunochemical assays. Nevertheless functional assays can be biased by the antithrombin III (AT III)-heparin complex activity; in fact trace amounts of heparin generally contaminate dermatan sulfate (DS) commercial preparations used as HC II reaction activators. We have employed a purified DS preparation showing these features: DS concentration 94.4%, chondroitin sulfate 5.6%, M.W. 21.4 kDa. The absence of any interference due to AT III-heparin complexes was verified in a kinetic HC II assay of some human plasma pools. The immunological inhibition of AT III by anti-AT III caused a minimal decrease (6-8%) of the reaction slope, attributable to AT III activity. Progressive increase of heparin concentration in the assay was effective only starting from 30 U/ml (the assay was carried out in the presence of polybrene to prevent any AT III activation). The reference interval (mean +/- SD) obtained from 157 normal subjects was 100.8 +/- 20.2%; there was a good correlation with immunoreactive HC II. The purified DS we have used seems suitable for routinary assays of HC II where a minimal interference due to AT III-heparin is required.
就免疫化学检测而言，一般更倾向于采用HC II功能检测。然而，功能检测可能会受到抗凝血酶III（AT III）-肝素复合物活性的影响；实际上，用作HC II反应激活剂的硫酸皮肤素（DS）商业制剂通常会被痕量肝素污染。我们使用了一种具有以下特性的纯化DS制剂：DS浓度94.4%，硫酸软骨素5.6%，分子量21.4 kDa。在对一些人血浆样本进行的HC II动力学检测中，证实不存在因AT III-肝素复合物导致的任何干扰。抗AT III对AT III的免疫抑制使反应斜率出现最小程度的下降（6 - 8%），这归因于AT III的活性。检测中肝素浓度的逐步增加仅在30 U/ml时才开始有效（检测在鱼精蛋白存在的情况下进行，以防止任何AT III激活）。从157名正常受试者获得的参考区间（均值±标准差）为100.8 ± 20.2%；与免疫反应性HC II有良好的相关性。我们所使用的纯化DS似乎适用于对AT III-肝素干扰最小的HC II常规检测。
