Vissac A M, Grimaux M, Chartier S, Chan F A, Chambrette B, Amiral J
Serbio Research Laboratory, Gennevilliers, France.
Thromb Res. 1995 May 15;78(4):341-52. doi: 10.1016/0049-3848(95)91462-t.
D.Dimer is currently used as a diagnotic help in thromboembolic events. The first application widely validated concerns the exclusion diagnosis of deep vein thrombosis and pulmonary embolism. In this context D.Dimer measurements must be performed individually and they must offer a good accuracy in evaluating the clinical decision threshold which is of 0.5 micrograms/ml when D.Dimer is expressed as initial fibrinogen equivalent. For this objective we report a new membrane based ELISA technique, which uses an immunofiltration device and two complementary monoclonal antibodies. The first one is coated onto the membrane and is used for the D.Dimer capture. The bound analyte is then revealed later using the second monoclonal antibody coupled to alkaline phosphatase. The assay is performed in less than 10 minutes and it can be used instantaneously by the clinical laboratories in emergency situations. Only 200 microliters of a standard citrated plasma are required. All samples containing more than 0.5 micrograms/ml D.Dimer produce a color development which intensity is a relation of the D.Dimer concentration. All specimen with levels below 0.3 micrograms/ml give negative tests, whereas a grey zone is present between 0.3 and 0.5 micrograms/ml. This assay offers all the specifications required by its applications to the exclusion diagnosis of deep vein thrombosis and pulmonary embolism.
D - 二聚体目前在血栓栓塞事件中用作诊断辅助手段。首次得到广泛验证的应用涉及深静脉血栓形成和肺栓塞的排除诊断。在此背景下，D - 二聚体检测必须单独进行，并且在评估临床决策阈值时必须具有良好的准确性，当D - 二聚体以初始纤维蛋白原当量表示时，该阈值为0.5微克/毫升。为实现这一目标，我们报告了一种基于膜的新型酶联免疫吸附测定（ELISA）技术，该技术使用免疫过滤装置和两种互补单克隆抗体。第一种抗体包被在膜上，用于捕获D - 二聚体。然后使用与碱性磷酸酶偶联的第二种单克隆抗体检测结合的分析物。该检测在不到10分钟内完成，临床实验室在紧急情况下可即时使用。仅需200微升标准枸橼酸盐血浆。所有D - 二聚体含量超过0.5微克/毫升的样本都会产生显色反应，其强度与D - 二聚体浓度相关。所有水平低于0.3微克/毫升的样本检测结果为阴性，而在0.3至0.5微克/毫升之间存在一个灰色区域。该检测提供了其用于深静脉血栓形成和肺栓塞排除诊断应用所需的所有规格。
