The present study was carried out for clinical evaluation of point-of-care-testing (POCT) of heart-type fatty acid-binding protein (H-FABP), Rapicheck H-FABP, for the diagnosis of acute myocardial infarction (AMI), in comparison with conventional cardiac biochemical markers such as myoglobin, creatine kinase isoenzyme MB (CK-MB), and cardiac troponin T. Whole blood samples from patients with confirmed AMI (n = 53), patients with non-AMI cardiac diseases (n = 24), and patients with non-cardiac diseases with chest pain (n = 6) were used. When a test line appeared within 15 min after the addition of 150 microL of whole blood, it was designated to be positive for H-FABP. A control line indicates a proper use of the test. On the other hand, when no test line appeared, it was negative. In the superacute phase of AMI within 3 hours, the diagnostic sensitivity of H-FABP was 93.1%, which was the highest of the four markers compared here. The diagnostic specificity in the phase for H-FABP was 64.3%, while it was 100% with cardiac troponin T. The POCT of H-FABP is thought to be practical for the detection of cardiac damage and effective for the diagnosis of AMI in superacute phase within 3 hours and/or 6 hours.