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Serum myoglobin levels were measured in normal subjects and patients by means of a newly developed radioimmunoassay. Myoglobin was identified in all of 135 sera from normal adults and ranged between 6 and 85 ng/ml (mean +/- SE 31 +/- 1.3). Raised myoglobin levels were present in 62 of 64 patients with documented acute myocardial infarction, the mean serum concentration being 528 +/- 76 ng/ml. Serial determinations in 46 patients with acute infarct showed that maximum values usually occurred within 4 hours after admission. In 19 of 42 cases, raised myoglobin levels preceded the rise in creatine kinase (CK) values; in the remaining patients, both serum myoglobin and creatine kinase were increased on admission. Only 2 of an additional 44 patients admitted with chest pain but without subsequent electrocardiographic, enzyme, or technetium-99m stannous pyrophosphate myocardial scintigraphic evidence of acute myocardial infarction had raised myoglobin levels; the mean value for this group was within the normal range (44 +/- 6 ng/ml). Serum myoglobin values also were normal in patients with congestive heart failure without acute myocardial infarction, and in patients after moderate exercise and cardiac catheterisation. Trasient myoglobinaemia appears to be one of the earliest laboratory abnormalities occurring in acute myocardial infarction and, therefore, should prove useful as a diagnostic aid in patients.