Introduction: T wave alternans (TWA) is a promising technique for detecting arrhythmia vulnerability. Previous studies in animals demonstrated that the magnitude of TWA is dependent on heart rate. However, the effects of heart rate on TWA in humans and the clinical relevance of this effect remain controversial. Methods ann Results: This was a prospective evaluation of pacing rate and monitoring lead configuration on TWA in subjects undergoing electrophysiologic study. Measurements of TWA mere performed on 45 patients in the absence of antiarrhythmic drugs. Recordings were made in normal sinus rhythm and during atrial pacing at 100 and 120 beats/min. Sustained monomorphic ventricular tachycardia (VT) was induced in 29 patients with programmed stimulation, TWA in the vector magnitude lead increased with heart rate, independent of VT inducibility (0.4 +/- 0.7 mu V, 1.6 +/- 1.9 mu V, and 2.4 +/- 2.1 mu V in sinus rhythm and at 100 and at 120 beats/min, respectively; P < 0.001). In addition, the diagnostic performance of TWA for inducible VT mas dependent on heart rate (sensitivity 4%, 42%, and 65%, and specificity 100%, 93%, and 63% at 77, 100, and 120 beats/min, respectively). By analyzing orthogonal Leads rather than the vector magnitude lead, the sensitivity is increased from 42% to 59% at 100 beats/min, but the specificity is reduced from 93 % to 72%. Conclusion: These results indicate that TWA in humans is strongly dependent on heart rate with regard to both magnitude and diagnostic performance, The optimal heart rate for the measurement of TWA is between 100 and 120 beats/min and multiple leads should be monitored.