In a multicenter study we compared three tests for ischemic myocardial injury (IMI): a new, automated enzyme immunoassay for S-troponin T (S-TNT; Boehringer Mannheim) and two S-creatine kinase (CK) isoenzyme MB assays (mass and catalytic concentrations). For critical evaluation of clinical sensitivity, we studied 243 cases with an IMI prevalence of 43% and an 18% prevalence of cases with unstable angina. Relative peak values of S-TNT and S-CK-MB (mass) after onset of pain were four- to fivefold higher than S-CK-MB (catalytic) results. Increases of S-TNT and S-CK-MB (mass), even though still within their reference ranges, indicated minor myocardial damage in about one-third of the cases primarily classified as unstable angina. The diagnostic window for S-TNT ranged from hours to weeks after the acute episode. The time courses were frequently biphasic, with the initial S-TNT peak closely paralleling that of the mass concentrations of S-CK-MB. With a biological half-life for S-TNT of 2 h, the prolonged increases in S-TNT indicate a continuous release of S-TNT from necrotizing cells. Clinical specificities of S-TNT and S-CK-MB (mass) were greater than that of S-CK-MB (catalytic), even in the presence of 30% to 40% severe skeletal muscle injuries. The combination of S-TNT and S-CK-MB (mass) is excellent for detection of acute IMI, including minor myocardial damage.