Hetland O, Goransson L, Nilsen D W
Department of Clinical Chemistry, Central Hospital of Rogaland, Stavanger, Norway.
Scand J Clin Lab Invest. 1995 Dec;55(8):701-13. doi: 10.3109/00365519509075400.
We have adapted the cardiac troponin T (TnT) immunoassay system (ELISA troponin T, Boehringer Mannheim, Germany), which is based upon streptavidin-biotin immunoassay technology, to a sensitive microplate system. A coating of microplates with biotinylated bovine serum albumin (biotin-LC-BSA) remained stable for months. A secondary streptavidin coating was prepared as the first step of the assay. By using o-phenylenediamine (o-PD) as a substrate for the peroxidase-anti-TnT conjugate, the system allowed rapid kinetic measurement of TnT levels. The upper limit of a reference population (97.5th percentile) was found to be 0.04 mu g l-1. Intra-assay imprecision at 0.08 mu g l-1 was 8%, and 3-4% between 0.28 and 4 mu g l-1. Between-assay imprecision was 6.2% at 0.28 mu g l-1. Studies of TnT and CK-MB mass concentration in acute myocardial infarction patients, treated with streptokinase, demonstrated a clinical sensitivity of the TnT microplate system similar to that of the CK-MB mass concentration test, during the first 8 h after initiation of thrombolytic therapy, at discriminator levels of 0.1 mu g l-1 (TnT) and 8 mu g l-1 (CK-MB mass concentration). The early CK-MB/TnT ratio was lower in patients with signs of successful reperfusion (early peak CK-MB) than in the remaining patients (p<0.001). Serum samples from two patients with renal failure and one patient with rhabdomyolysis demonstrated strong non-linear behaviour with dilution, indicating the presence of an interfering factor. Kinetic measurement compared favourably with end-point analysis with respect to sensitivity and total analysis time. The system described greatly reduces the costs of TnT measurements compared to the ES systems. The total assay time is 70-90 min.
我们已将基于链霉亲和素-生物素免疫测定技术的心肌肌钙蛋白T（TnT）免疫测定系统（酶联免疫吸附测定法TnT，德国宝灵曼公司）应用于灵敏的微孔板系统。用生物素化牛血清白蛋白（生物素-LC-BSA）包被微孔板，其稳定性可维持数月。作为测定的第一步，制备二级链霉亲和素包被。通过使用邻苯二胺（o-PD）作为过氧化物酶-抗TnT缀合物的底物，该系统可快速动态测定TnT水平。参考人群的上限（第97.5百分位数）为0.04μg l-1。在0.08μg l-1时，批内不精密度为8%，在0.28至4μg l-1之间为3-4%。在0.28μg l-1时，批间不精密度为6.2%。对接受链激酶治疗的急性心肌梗死患者的TnT和CK-MB质量浓度进行研究，结果表明，在溶栓治疗开始后的前8小时内，当鉴别水平为0.1μg l-1（TnT）和8μg l-1（CK-MB质量浓度）时，TnT微孔板系统的临床敏感性与CK-MB质量浓度检测相似。成功再灌注（早期CK-MB峰值）患者的早期CK-MB/TnT比值低于其余患者（p<0.001）。两名肾衰竭患者和一名横纹肌溶解患者的血清样本在稀释时表现出强烈的非线性行为，表明存在干扰因素。在灵敏度和总分析时间方面，动态测定优于终点分析。与酶标仪系统相比，所述系统大大降低了TnT测量成本。总测定时间为70-90分钟。
