DEVELOPMENT AND CHARACTERIZATION OF A RAPID ASSAY FOR BEDSIDE DETERMINATIONS OF CARDIAC TROPONIN-T

Background The appearance of cardiac proteins in blood is the most spe cific and sensitive indicator of acute myocardial cell necrosis. The m easurement of cardiac markers, however, is time consuming and requires sophisticated equipment. To facilitate the biochemical detection for acute myocardial cell necrosis, a whole-blood rapid assay device for c ardiac troponin T detection was developed that provides a test result within 20 minutes. Methods and Results Monoclonal antibody M7 is label ed with gold particles, and antibody 1B10 is labeled with biotin. Both antibodies, as well as buffer substances and detergents, are adsorbed onto paper fleeces mounted below an application well. Heparinized blo od (160 mu L) applied to this well solubilizes the dry chemistry reage nts. Blood cells are separated from plasma via a glass-fiber fleece. T he immunocomplexes formed are concentrated within the reading zone by binding of the biotin-labeled antibody with streptavidine immobilized to the test device. Troponin T bound to the test device serves as a co ntrol. The detection limit of this assay is 0.18 mu g/L with a cross-r eactivity with skeletal troponin T of 0.5%. In clinical analyses invol ving 25 healthy volunteers, 62 patients with chest pain but without my ocardial ischemia, 35 patients with acute myocardial infarction, 24 pa tients with minor myocardial cell damage due to radiofrequency ablatio n, and 35 patients with unstable angina, the rapid assay was comparabl e to the troponin T enzyme immunoassay in regard to sensitivity and sp ecificity. Conclusions This newly developed assay allows accurate, rap id, and convenient diagnosis of acute myocardial cell necrosis.